US20130258075A1 - Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles - Google Patents
Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles Download PDFInfo
- Publication number
- US20130258075A1 US20130258075A1 US13/788,165 US201313788165A US2013258075A1 US 20130258075 A1 US20130258075 A1 US 20130258075A1 US 201313788165 A US201313788165 A US 201313788165A US 2013258075 A1 US2013258075 A1 US 2013258075A1
- Authority
- US
- United States
- Prior art keywords
- stream
- ejected
- flow cytometer
- trajectory
- micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1404—Fluid conditioning in flow cytometers, e.g. flow cells; Supply; Control of flow
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
- G01N2015/144—Imaging characterised by its optical setup
Definitions
- the present technology relates to a micro-particle sorting apparatus and a method of determining a trajectory of an ejected stream of the micro-particle sorting apparatus.
- the present technology relates to a micro-particle sorting apparatus which automatically determines a trajectory of a fluid stream or the like carrying micro-particles that is ejected from an orifice.
- micro-particle sorting apparatus for example, a flow cytometer
- detects characteristics of micro-particles such as cells optically, electrically or magnetically, and fractionates or sorts the micro-particles so as to collect only micro-particles having predetermined characteristics.
- a fluid stream (laminar flow of sample liquid and sheath liquid containing cells) is generated from an orifice formed in a flow cell, a vibration is applied to the orifice to transform the fluid stream into a form of liquid droplets, and electric charge is applied to the liquid droplets. Then, the movement direction of the liquid droplets containing the micro-particles discharged from the orifice is electrically controlled to collect target micro-particles having the desired characteristics and non-target micro-particles having characteristics other than those desired are sorted into separate collection containers.
- a micro-particle sorting apparatus including: a microchip in which a flow path through which liquid containing micro-particles flows and an orifice through which the liquid flowing through the flow path is discharged into a space outside the chip are disposed; a vibrating element configured to transform the liquid into a form of liquid droplets and discharge the liquid droplets in the orifice; charge means for adding an electric charge to the discharged liquid droplets; optical detection means for detecting the optical characteristics of micro-particles which flow through the flow path; a pair of electrodes provided so as to be opposed to each other with the moving liquid droplets interposed therebetween along a movement direction of the liquid droplets discharged into the space outside the chip; and two or more containers that collect the liquid droplets passing through between the pair of electrodes”.
- a micro-particle sorting apparatus it is desirable to design fluid collection such that a fluid stream or liquid droplets generated from an orifice formed in a flow cell or a microchip enter inside a collection container. Accordingly, it is necessary to prevent a deviation of the fluid stream or the liquid droplets from an assumed direction.
- the prevention of the deviation has been performed by checking an ejected fluid stream or the like with visual observation of a user, and problems regarding reliability and stability were generated and depended upon the experience level of the user. Further, performing the determination of the deviation with visual observation was extremely complicated in the configuration of the apparatus.
- micro-particle sorting apparatus capable of automatically detecting a deviation of a trajectory of an ejected fluid stream or liquid droplets that carries the micro-particles.
- micro-particles broadly includes biologically-relevant micro-particles such as cells, microorganisms, liposomes, and the like, and synthetic particles such as latex particles, gel particles, industrial particles, and the like.
- the biologically-relevant micro-particles include chromosomes, liposomes, mitochondria, organelles, and the like configuring various cells.
- Cells include animal cells (hematopoietic cells and the like) and plant cells.
- Microorganisms include bacteria such as coli bacteria, viruses such as tobacco mosaic virus, and fungi such as yeast cells.
- Biologically-relevant micro-particles include nucleic acids, proteins, and biologically-relevant macromolecules such as a complex thereof.
- Industrial particles may be organic or inorganic high polymer materials, metals and the like, for example.
- Organic polymer materials include polystyrene, styrene-divinylbenzene, polymethylmethacrylate and the like.
- Inorganic polymer materials include glasses, silica, magnetic materials, and the like.
- Metals include gold colloids, aluminum and the like.
- the shape of the micro-particles is generally spherical, however it may be non-spherical and the size, weight and the like thereof are not particularly limited.
- a flow cytometer comprises a micro-orifice configured to eject a fluidic stream, and an imaging device configured to image an ejected stream, wherein the ejected stream is at least a portion of the fluidic stream ejected from the micro-orifice.
- the flow cytometer may further include at least one processor that is configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image.
- the ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets.
- the micro-orifice may be an exit orifice of a micro-fluidic chip.
- the flow cytometer may further comprise electrostatic deflection apparatus configured to deflect the liquid droplets, and charging apparatus configured to apply charge to individual liquid droplets so that individual liquid droplets carrying micro-particles may be sorted according to pre-selected sorting criteria.
- the flow cytometer may further comprise automated focusing apparatus, wherein the at least one processor is further configured to measure a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and control the focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image. The focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- the at least one processor may be configured to evaluate the trajectory of the ejected stream based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream.
- the at least one processor may be configured to evaluate the trajectory by identifying a line that connects the one or more contrasted spots.
- the at least one processor may be configured to compute an angle associated with the trajectory of the ejected stream.
- the computed angle may be a measure of deviation of the ejected stream from a predetermined direction.
- the at least one processor may be further configured to detect an abnormality in operation of the flow cytometer if the computed angle is greater than a predetermined threshold value.
- the detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice.
- the at least one processor may be further configured to execute an alerting function if the computed angle is greater than a predetermined threshold value.
- the flow cytometer may further comprise movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- the at least one processor may be configured to calculate the trajectory of the ejected stream in a focus direction based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream.
- the first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream.
- the at least one processor may be configured to calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- the at least one processor may be further configured to identify a width of the ejected stream in the received image and determine a diameter of the micro-orifice based upon the identified width of the ejected stream.
- Embodiments also include a trajectory evaluation system for a flow cytometer.
- the trajectory evaluation system may comprise an imaging device configured to image the ejected stream, wherein the ejected stream is at least at portion of a fluidic stream ejected from a micro-orifice of the flow cytometer.
- the trajectory evaluation system may further comprise at least one processor configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image.
- the ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets.
- the micro-orifice may be an exit orifice of a micro-fluidic chip.
- the trajectory evaluation system may include automated focusing apparatus, wherein the at least one processor is further configured to measure a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and control the focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image.
- the focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- the at least one processor may be configured to evaluate the trajectory of the ejected stream based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream.
- the at least one processor may be configured to evaluate the trajectory by identifying a line that connects the one or more contrasted spots.
- the at least one processor may be configured to compute an angle associated with the trajectory of the ejected stream.
- the computed angle may be a measure of deviation of the ejected stream from a predetermined direction.
- the at least one processor may be further configured to detect an abnormality in operation of a flow cytometer if the computed angle is greater than a predetermined threshold value.
- the detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice.
- the at least one processor may be further configured to execute an alerting function if the computed angle is greater than a predetermined threshold value.
- the trajectory evaluation system may provide a signal for moving movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- the at least one processor may be configured to calculate the trajectory of the ejected stream in a focus direction based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream.
- the first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream.
- the at least one processor may be configured to calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- the at least one processor may be further configured to identify a width of the ejected stream in the received image and determine a diameter of the micro-orifice based upon the identified width of the ejected stream.
- trajectory evaluation system may be implemented in a trajectory evaluation system for a flow cytometer in any combination.
- Embodiments also include a method of measuring a trajectory of an ejected stream in a flow cytometer.
- the method may comprise an act of imaging, with an imaging device, the ejected stream, wherein the ejected stream is at least a portion of a fluidic stream ejected from a micro-orifice of the flow cytometer.
- the method may further include receiving, by at least one processor, an image of the ejected stream imaged by the imaging device, and processing, by the at least one processor, the received image to detect one or more contrasted spots located centrally within the ejected stream and to evaluate a trajectory of the ejected stream.
- the ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets.
- the micro-orifice may be an exit orifice of a micro-fluidic chip.
- the method of measuring a trajectory may include an act of automated focusing, wherein the at least one processor measures a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and controls focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image.
- the focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- the method of measuring a trajectory may comprise evaluating the trajectory of the ejected stream, by the at least one processor, based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream.
- the at least one processor may evaluate the trajectory by identifying a line that connects the one or more contrasted spots.
- the at least one processor may compute an angle associated with the trajectory of the ejected stream.
- the computed angle may be a measure of deviation of the ejected stream from a predetermined direction.
- the at least one processor may detect an abnormality in operation of a flow cytometer if the computed angle is greater than a predetermined threshold value.
- the detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice.
- the at least one processor may execute an alerting function if the computed angle is greater than a predetermined threshold value.
- the method of measuring a trajectory may include providing a signal, by the at least one processor, for moving movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- the method of measuring a trajectory may include calculating the trajectory of the ejected stream in a focus direction, by the at least one processor, based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream.
- the first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream.
- the at least one processor may calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- the method of measuring a trajectory may further comprise identifying, by the at least one processor, a width of the ejected stream in the received image and determining a diameter of the micro-orifice based upon the identified width of the ejected stream.
- FIG. 1 is a schematic diagram illustrating a configuration of a sorting system of a micro-particle sorting apparatus (flow cytometer) according to an embodiment of the present technology.
- the flow cytometer may be configured as a microchip flow cytometer.
- FIGS. 2A and 2B are schematic diagrams illustrating a configuration of an example of a microchip which is mountable on a flow cytometer.
- FIGS. 3A to 3C are schematic diagrams illustrating a configuration of an orifice of a microchip.
- FIG. 4 is a flowchart illustrating steps for determining a trajectory of a fluid stream or the like of a flow cytometer.
- FIGS. 5A and 5B are pictures showing an example of images before and after focusing which are imaged by a droplet camera of a flow cytometer.
- FIGS. 6A and 6B are schematic diagrams showing images of fluid streams having different widths from each other imaged by a droplet camera of a flow cytometer.
- FIGS. 7A and 7B are pictures showing an example of images of a fluid stream and liquid droplets imaged by a droplet camera of a flow cytometer.
- FIGS. 8A and 8B are pictures showing an example of images of a fluid stream and liquid droplets imaged by a droplet camera of a flow cytometer.
- FIGS. 9A and 9B are pictures showing an example of images of liquid droplets imaged by a droplet camera of a flow cytometer.
- FIGS. 10A and 10B are schematic diagrams showing an example of images of fluid streams imaged by a droplet camera of a flow cytometer.
- FIG. 1 is a schematic diagram illustrating a configuration of a sorting system of a micro-particle sorting apparatus 1 (hereinafter, also referred to as a “flow cytometer 1 ”) according to an embodiment of the present technology.
- the flow cytometer is configured as a microchip flow cytometer.
- Reference numeral 11 in the drawing denotes a chip loading module storing a microchip 2 .
- the chip loading module 11 includes a chip loading unit which performs transportation to store the microchip 2 inserted from the outside to a predetermined position, and a transporting liquid connecting unit which supplies sample liquid, sheath liquid, and the like including micro-particles to the stored microchip 2 (both not shown).
- the chip loading module 11 includes a chip vibrating unit which is formed in the microchip 2 , and applies a vibration to the orifice 21 generating laminar flow (flow stream S) of sample liquid and sheath liquid to transform the fluid stream S into a form of liquid droplets and discharge the liquid droplets, and an electric charge unit which applies an electric charge to the discharged liquid droplets (both not shown).
- FIGS. 2A to 3C show an example of the microchip 2 which is mountable on the flow cytometer 1 .
- FIG. 2A shows a schematic diagram of the upper surface and FIG. 2B shows a cross-sectional schematic diagram taken along the line IIB-IIB of FIG. 2A .
- FIGS. 3A to 3C are diagrams schematically illustrating one configuration of the orifice 21 of the microchip 2 .
- FIG. 3A shows a schematic diagram of the upper surface
- FIG. 3B shows a cross-sectional schematic diagram
- FIG. 3C shows a plan diagram.
- FIG. 3B is a cross-sectional diagram taken along the line IIIB-IIIB of FIG. 2A .
- the microchip 2 may be formed by bonding substrate layers 2 a and 2 b on which a sample flow path 22 is formed.
- the sample flow path 22 on the substrate layers 2 a and 2 b can be formed by injection molding of thermoplastic resin using mold.
- thermoplastic resin existing plastics of the related art as materials of the microchip such as polycarbonate, polymethylmethacrylate resin (PMMA), cyclic polyolefins, polyethylene, polystyrene, polypropylene, polydimethylsiloxane (PDMS) or the like may be used.
- the sample liquid is introduced to a sample inlet 23 from the transporting liquid connecting unit and joins with the sheath liquid which is introduced to a sheath inlet 24 from the transporting liquid connecting unit to transport the liquid to the sample flow path 22 .
- the sheath liquid introduced from the sheath inlet 24 is transported by dividing into two directions. Then, in the joining portion with the sample liquid introduced from the sample inlet 23 , the sheath liquid joins with the sample liquid so as to interpose the sample liquid from two directions. Accordingly, in the joining portion, three-dimensional laminar flow is formed in which the laminar flow of the sample liquid is positioned at the center of the laminar flow of the sheath liquid.
- Reference numeral 25 denotes a suction flow path for removing clogging and bubbles by causing a negative pressure inside the sample flow path 22 to counterflow temporarily, when clogging and bubbles are generated in the sample flow path 22 .
- a suction outlet 251 which is connected to a negative pressure source such as a vacuum pump or the like through the transporting liquid connecting unit is formed, and another end thereof is connected to the sample flow path 22 in a communicating port 252 .
- the laminar flow width of the three-dimensional laminar flow may be formed to be narrowed down in narrowing units 261 (see FIGS. 2A and 2B ) and 262 (see FIGS. 3A to 3 C) so that the area of the perpendicular cross section with respect to the transporting liquid direction becomes small gradually or in steps from the upstream to the downstream in the transporting liquid direction.
- the three-dimensional laminar flow becomes the fluid stream S (see FIG. 1 ) and is discharged from the orifice 21 provided at one end of the flow path.
- the discharging direction of the fluid stream S from the orifice 21 is shown as the positive Y axis direction.
- the characteristics of the micro-particles may be detected between the narrowing unit 261 and the narrowing unit 262 of the sample flow path 22 .
- a light irradiation detecting unit now shown, a laser is emitted with respect to the micro-particles which are arranged in a line in the center of the three-dimensional laminar flow to flow inside the sample flow path 22 , and scattering light and fluorescence generated from the micro-particles are detected by one or more light detectors.
- a connecting unit of the sample flow path 22 and the orifice 21 is set as a straight unit 27 formed to be linear.
- the straight unit 27 functions for ejecting the fluid stream S from the orifice 21 linearly in the positive Y axis direction.
- the fluid stream S ejected from the orifice 21 may be transformed into a form of liquid droplets by the vibration applied to the orifice 21 by a chip vibrating unit.
- the orifice 21 is opened in the end surface direction of the substrate layers 2 a and 2 b , and a cut-out portion 211 is provided between the opening position and the end surface of the substrate layers.
- the cut-out portion 211 is formed by cutting out the substrate layers 2 a and 2 b between the opening position of the orifice 21 and the end surface of the substrates so that a diameter L of the cut-out portion 221 is larger than a diameter 1 of the opening of the orifice 21 (see FIG. 3C ). It is desirable that the diameter L of the cut-out portion 211 be formed to be larger by more than double the diameter 1 of the opening of the orifice 21 so as not to interrupt the movement of the liquid droplets discharged from the orifice 21 .
- Reference numerals 12 and 12 in FIG. 1 denote a pair of deflection plates which are arranged to oppose each other by interposing the fluid stream S (or the discharged liquid droplets) which is ejected from the orifice 21 and imaged by a droplet camera 4 which will be described later.
- the deflection plates 12 and 12 include electrodes which control the movement direction of the liquid droplets discharged from the orifice 21 by an electric force interacting with electric charge applied to the liquid droplets.
- the deflection plates 12 and 12 also control the trajectory of the fluid stream S generated from the orifice 21 by an electric force interacting with electric charge applied to the fluid stream S.
- the opposing direction of the deflection plates 12 and 12 is shown as the X axis direction.
- the fluid stream S (or liquid droplets D thereof) may be collected in any of a plurality of collection tubes (collection containers) 3 which are arranged in a line in the opposing direction (X axis direction) of the deflection plates 12 and 12 (see FIG. 1 ).
- the collection tubes 3 may be general-purpose plastic tubes or experimental glass tubes.
- the number of the collection tubes 3 is not particularly limited, but the embodiment shows a case of arranging five collection tubes.
- the fluid stream S generated from the orifice 21 is introduced to any one of the five collection tubes 3 depending on the existence or non-existence, or the size of the electric force acting between the deflection plates 12 and 12 and collected therein.
- the collection tubes 3 may be disposed in a collection tube container 31 in an exchangeable manner.
- the collection tubes 3 are disposed in the movement direction (X axis direction) shown as an arrow F 1 in FIG. 1 in a movable manner.
- the collection tubes 3 may be disposed so that only the collection tubes 3 move in the X axis direction in a state where the collection tube container 31 is fixed, or the collection tubes 3 may be disposed movably with the movement of the collection tube container 31 .
- the collection tube container 31 may be disposed on a Z axis stage 32 which is configured to be movable in a direction (Z axis direction) perpendicular to the discharging direction (Y axis direction) of the fluid stream S from the orifice 21 and the opposing direction (X axis direction) of the deflection plates 12 and 12 .
- An arrow F 2 in FIG. 1 denotes the movement direction of the Z axis stage 32 .
- Reference numeral 321 in the drawing denotes a waste liquid port provided on the Z axis stage 32 .
- the collection tube container 31 and the Z axis stage 32 configure a collection unit 33 which is driven by a Z axis motor (not shown).
- a droplet camera 4 may be any suitable camera (CCD camera, CMOS image sensor or the like) for imaging the fluid stream S ejected from the orifice 21 of the microchip 2 or the liquid droplets discharged therefrom (see FIG. 1 ).
- the droplet camera 4 may be designed to be able to perform automated focusing under the control of at least one processor on the captured image of the fluid stream S or the liquid droplets.
- the image captured by the droplet camera 4 may be displayed on the display unit such as a display, and used for a user to check for the formation state (size, shape, intervals and the like of the liquid droplets) of the liquid droplets of the orifice 21 .
- the trajectories of the fluid stream S (or liquid droplets) ejected from the orifice 21 are different depending on the individual differences of the mounted microchips 2 , and the position of the fluid stream S can be changed in the Z axis direction (and X axis direction) in the drawing, at each time of exchanging the microchip 2 .
- Continuing ejecting the fluid stream S or continuing discharging the liquid droplets may result in the degradation or the like of the microchip 2 , so that the position of the fluid stream S (or the liquid droplets) can be changed over time in the Z axis direction (and X axis direction) in the drawing.
- the droplet camera 4 also functions for detecting such position change of the fluid stream S (or the liquid droplets) in the Z axis direction (and X axis direction).
- the flow cytometer 1 includes a light irradiation detecting unit for detecting the optical characteristics of micro-particles, a data analysis unit for determining the characteristics, a tank unit which stores the sample liquid and the sheath liquid, and a control unit for controlling each configuration thereof, which are included in general flow cytometers.
- the control unit may be configured by a general-purpose computer including at least one CPU, a memory or a hard disk and the like, and an OS.
- Machine-readable instructions that may be executed by the at least one CPU may be stored in memory and, when executed by the at least one CPU, specially adapt the computer for executing each step of the position control, which will be described later, and other processes of the flow cytometer.
- the light irradiation detecting unit may be configured by a laser light source, an irradiation system which includes a condensing lens, a dichroic minor, a bandpass filter and the like which condense and emit the laser with respect to the micro-particles, and a detecting system which detects the measuring target light generated from the micro-particles by excitation of the laser.
- the detecting system may be configured by an area imaging device or the like such as a PMT (photomultiplier tube), a CCD, or a CMOS element.
- the measuring target light which is detected by the detecting system of the light irradiation detecting unit is the light which is generated from the micro-particles by the emission of the measuring light, and can be scattered light such as forward-scattered light, backward-scattered light, Rayleigh-scattered or Mie scattered light, or fluorescence.
- the above measuring target light is converted into electrical signals, output to the control unit and provided for determining the optical characteristics of the micro-particles.
- the flow cytometer 1 may magnetically or electrically detect the characteristics of the micro-particles.
- microelectrodes are arranged to oppose each other in the sample flow path 22 of the microchip 2 , and a resistance value, a capacitance value, an inductance value, impedance, a changing value of the electric field between the electrodes, or the change in magnetization, magnetic field, and the like are measured.
- FIG. 4 is a flowchart illustrating steps for determining the trajectory of the fluid stream S (or the liquid droplets) of the flow cytometer 1 , according to one embodiment.
- the steps for determining the trajectory include processes of a “fluid stream generating step S 1 ,” a “droplet camera Z axis scanning and fluid stream imaging step S 2 ,” a “focusing step S 3 ,” a “center line detecting step S 4 ,” a “displaying step S 5 ,” an “orbital direction determining step S 6 ,” an “alerting step S 7 ,” and a “collection tube moving and aligning step S 8 .”
- each process will be described.
- the transporting liquid connecting unit starts transporting the sample liquid and the sheath liquid to the sample inlet 23 and the sheath inlet 24 of the microchip 2 , and a fluid stream S is ejected from the orifice 21 (see FIG. 4 ).
- the control unit outputs the signals to the transporting liquid connecting unit and starts transporting the sample liquid and the sheath liquid.
- the fluid stream S ejected from the orifice 21 may be collected in the waste liquid port 321 and disposed of.
- the chip vibrating unit applies the vibration to the orifice 21 , and the liquid droplets may be discharged instead of a continuous fluid stream S from the orifice, so that the liquid droplets can be collected in the waste liquid port 321 and disposed of.
- the control unit outputs the signals to the droplet camera 4 and the droplet camera 4 which receives the signals may be moved in the Z axis direction (see FIG. 4 ), for example, to center an image of the stream. Then, the control unit outputs the signals to the droplet camera 4 , and the droplet camera 4 which receives the signals performs imaging of the fluid stream S (or the liquid droplets).
- the focusing may be performed in the X axis direction when imaging the image of the fluid stream S (or the liquid droplets) by the droplet camera 4 (see FIG. 4 ).
- the image of the fluid stream S (or the liquid droplets) imaged by the droplet camera 4 may be output to the control unit, and the control unit may perform focusing control until detecting the contrasted or bright points in the image in the focusing step S 3 .
- the bright points denote one or a plurality of pixels having higher brightness than a predetermined threshold value in the image of the fluid stream S (or the liquid droplets) imaged by the droplet camera 4 .
- a contrasted point or spot may be a spot having a luminance or color significantly different (e.g., greater than about 10% variation) from a background luminance or color around the spot.
- a contrasted spot may be a gray spot on a white background, a yellow spot on a red background, a white spot on a black background, etc. in a recorded image.
- FIG. 5A represents a picture showing an example of a state before the focusing of the imaged liquid droplets is performed (see FIG. 5A )
- FIG. 5B represents a picture showing an example of a state after the focusing of the liquid droplets is performed (see FIG. 5B ).
- FIG. 5B since the focusing of the image P is performed, it is possible to detect at least one bright point B in the center position of each liquid droplet D. Even in a case where the fluid stream S is ejected instead of the liquid droplets D from the orifice, it is possible to detect at least one bright point B in the center portion along the trajectory of the fluid stream S in the same manner.
- the control unit can determine whether the image P is in a focused state.
- control unit may determine the diameter of the orifice, based on the width of the fluid stream S detected in the direction perpendicular (Z axis direction) to the trajectory of the fluid stream S in the captured image P.
- FIGS. 6A and 6B show schematic diagrams of two captured images which have a different width of the fluid stream S from each other ( FIGS. 6A and 6B ).
- the control unit may be configured to determine accurately that the diameter of the orifice is 100 ⁇ m or the like, for example, by evaluating the width of the fluid stream S shown in FIG. 6A based on information stored in a memory unit.
- the control unit may be configured to determine accurately that the diameter of the orifice is 70 ⁇ m or the like, for example, by evaluating the width of the fluid stream S based on the information stored in the memory unit.
- the control unit may record or display the determined diameter of the orifice as the diameter of the orifice of the chip used in the flow cytometer 1 . Accordingly, a manual setting or recording of the diameter of the orifice by a user is not necessary, and thus it is possible to prevent setting mistakes such as mis-setting or mis-recording the diameter of the orifice.
- control unit may detect a center line of the fluid stream S from one or more bright points in the image of the fluid stream S (or the liquid droplets D) imaged by the droplet camera 4 , and may compare preset center line information with the detected center line (see FIG. 4 ).
- FIGS. 7A and 7B show states when a center line L of the fluid stream S (or the liquid droplets D) is detected in the captured image.
- the control unit may be configured to detect the straight line formed by the plurality of bright points displayed along the ejecting direction of the fluid stream S in the image of the fluid stream S imaged by the droplet camera 4 as the center line L.
- the control unit may identify the bright points B in the captured image P of the fluid stream S as the center line L.
- the control unit may be configured to detect a straight line formed by connecting one or more bright points displayed in each of the liquid droplets D as the center line L.
- the control unit may identify a line formed by connecting the bright points of each of the liquid droplets as the center line L of the liquid droplets.
- the control unit may set a line which most closely approximates the center line information which will be described later, as the center line.
- control unit can display the captured image on a display unit such as a display monitor (see FIG. 4 ).
- the control unit can arrange and display the fluid stream S (see FIG. 7A ) or the liquid droplets D (see FIG. 7B ) of the captured image in the center of such a display based on the center line L which is described above.
- the control unit may align the droplet camera 4 in the Z axis direction.
- the control unit may perform alignment based on the captured image P, until the number of the pixels of the positive direction side and the negative direction side of the Z axis direction become the same by setting the center line L as the boundary between positive side and negative side pixels.
- the image P of the fluid stream S (or the liquid droplets D) can be automatically aligned and displayed in the center of the display.
- the control unit may determine the trajectory of the fluid stream S (or the liquid droplets D) (see FIG. 4 ). In more detail, the control unit may determines a deviation of the trajectory in the Z axis direction and also a deviation of the trajectory in the X axis direction.
- the processes of “Z axis direction determining step S 61 ” and “X axis direction determining step S 62 ” are included. Each process will be described later.
- control unit may determine a trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction.
- the control unit may compare the center line L and predetermined center line information stored in the memory unit in advance. With respect to the fluid stream S (or the liquid droplets D), the center line L is detected as described above.
- the predetermined center line information may be information representing a straight line perpendicular to XZ plane stored in the memory unit in advance, and may further represent a line which makes the number of pixels of the positive direction side and the negative direction side of the Z axis direction the same by setting the predetermined center line as the boundary, in the captured image.
- FIGS. 8A and 8B also show states where the determined center line L of the fluid stream S (or the liquid droplets D) is detected in the captured image in the same manner as FIGS. 7A and 7B .
- the center line L is deviated by ⁇ 1 degrees in the YZ plane with respect to the predetermined center line information I (see FIG. 8A ).
- the center line is deviated by ⁇ 2 degrees in the YZ plane when compared to the predetermine center line information I (see FIG. 8B ).
- the process of the display step S 5 for making the number of the pixels of the positive direction side and the negative direction side of the Z axis direction the same by setting the center line L as the boundary by the control unit is omitted.
- control unit may determine that there is nearly no deviation of the center line L in the YZ plane with respect to the predetermined center line information I (see FIGS. 7A and 7B ).
- the control unit may also be configured to determine that an inclination angle (e.g., the angles ⁇ 1, or ⁇ 2) with respect to the center line information I of the center line L detected based on the comparison of the center line L and the center line information I, exceeds a predetermined threshold value, and determine that the microchip is abnormal.
- the control unit can determine the deviation of the trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction by comparing the center line information I and the center line L, and when the trajectory is deviated, the control unit can automatically determine that the microchip or the like is in a malfunction state (abnormal state of clogging or the like).
- An inclination angle that may result in an abnormal determination may be an inclination angle greater than 0.5 degree in some embodiments, greater than 1 degree in some embodiments, greater than 2 degrees in some embodiments, greater than 5 degrees in some embodiments, greater than 10 degrees in some embodiments, or greater than 20 degrees in some embodiments.
- An abnormal inclination angle may be an angle at which the ejected stream will no longer be captured by a collection vessel.
- control unit may determine a trajectory of the fluid stream S (or the liquid droplets D) in the X axis direction (see FIG. 4 ).
- FIGS. 9A and 9B show pictures of an example of the images of the liquid droplets in which the focusing may be performed.
- FIG. 9A when the trajectory of the liquid droplets D is not deviated in the X axis direction, since the focusing of the droplet camera 4 is performed based on the signals of the control unit, a focused region R 1 is detected for the length of the stream in the image, while a non-focused region R 2 is not detected.
- the control unit Since the control unit detects both the non-focused region R 2 and the focused region R 1 in the image P, the abnormity of the microchip or the like can be determined. Accordingly, when identifying the existence of the non-focused region R 2 in the image of the fluid stream S or the liquid droplets D and confirming the existence of the non-focused region R 2 , the control unit determines that the trajectory of the fluid stream S or the liquid droplets D is deviated in the X axis direction. Therefore, in the flow cytometer 1 , when the trajectory is deviated in the X axis direction, the control unit may automatically determine that the microchip or the like is in a malfunction state (abnormity state of clogging or the like).
- FIGS. 10A and 10B show schematic diagrams of the image of fluid stream S in which at least one bright point is detected.
- the control unit may perform focusing on a negative direction side of the Y axis direction, and at least one bright point B may be detected (see FIG. 10A ). Further, the control unit may perform focusing on a positive direction side of the Y axis direction, and at least one bright point B may be detected at a second focus position different from a first focus position found for the at least one bright point B shown in FIG. 10A (see FIG. 10B ).
- control unit can perform the focusing on two portions of the end portions (end portion of the positive Y axis direction side and the end portion of the negative Y direction side) of the fluid stream S in the image P of the fluid stream S.
- control unit can obtain position information corresponding to the deviation of the trajectory in the X axis direction, and in the collection tube moving and aligning step S 8 which will be described later, the aligning of the collection tubes 3 in the X axis direction can be performed by using the detected position information.
- the control unit may determine that the inclination or deviation angle exceeds a predetermined threshold value. In response, the control unit may perform alerting with respect to a user (see FIG. 4 ).
- various methods such as, a method for displaying a light or a message by a display unit such as a display, or a method for providing an output unit in the flow cytometer 1 and alerting by an audio output or the like, can be used as a method for alerting a user.
- the user can check for the, malfunction, breakage, or the like of the chip.
- the control unit may perform positioning of the collection tubes 3 based on the position information corresponding to the deviation of the trajectory in the X axis direction described above (see FIG. 4 ).
- the information regarding the trajectory of the fluid stream S (or the liquid droplets D) in the X axis direction is converted into the position information of the collection tubes 3 in the same direction, and the collection tubes 3 are moved to the position corresponding to the converted position information. Accordingly, the collection tubes 3 disposed in the collection tube container 31 and the fluid stream S are aligned in the X axis direction, and it is possible for the ejected fluid stream S to reach the collection tubes 3 precisely.
- the control unit may perform positioning of the collection tube container 31 based on the position information obtained by the aligning of the Z axis direction described above.
- the information regarding the trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction is converted into the position information of the collection tubes 3 in the same direction, and the Z axis stage 32 is moved to the position corresponding to the converted position information.
- the collection tubes 3 disposed in the collection tube container 31 and the fluid stream S are aligned in the Z axis direction, and it is possible for the ejected fluid stream S to reach the collection tubes 3 precisely.
- each process of the steps S 1 to S 8 has been described in order, however, the present technology is not limited to be executed in this order.
- the process of step S 7 may be executed after the process of the step S 8 .
- not all steps may be implemented.
- one or more steps may be repeated.
- a micro-particle sorting apparatus comprises an imaging device that images a fluid stream ejected from an orifice, or liquid droplets discharged from the orifice, and a control unit.
- the control unit may be configured to detects a center line of the fluid stream or the plurality of the liquid droplets from contrasted points in an image of the fluid stream or the liquid droplets imaged by the imaging device, and compare the center line with preset center line information.
- the micro-particle sorting apparatus may further include a display unit that displays the image.
- the imaging device may be configured to focus the captured image, and the control unit may performs focusing on at least a part of regions of the image.
- the control unit may be configured to determine, based upon a contrast ratio of selected portion of the image falling in a predetermined range, that the image is in a focused state or a non-focused state.
- the micro-particle sorting apparatus may comprise a microchip flow cytometer in which the orifice is provided in a microchip.
- control unit may be configured to identify or set a straight line corresponding to a plurality of the contrasted points of the fluid stream displayed along the ejection direction in the image of the fluid stream imaged by the imaging device.
- the straight line may be identified as the center line and trajectory of the ejected fluid stream from the orifice.
- control unit may be configured to identify or set the straight line corresponding to a plurality of the contrasted points of the liquid droplets displayed along the ejection direction discharged from the orifice in the image of the fluid stream imaged by the imaging device, and to identify the straight line as a center line and trajectory of the ejected droplets.
- the control unit may be configured to determine an abnormity of the ejected fluid stream or liquid droplets by calculating an inclination value between the identified center line and a predetermined reference line.
- the abnormality may be determined when a comparison between the center line and the reference line exceeds a predetermined threshold value.
- control unit may be configured to determine an existence of a non-focused region in the image of the fluid stream or the plurality of the liquid droplets.
- the control unit may determine an abnormity in the image when the non-focused region and a focused region are detected in the image of the fluid stream or the plurality of the liquid droplets.
- the micro-particle sorting apparatus may comprise a pair of deflection plates that are disposed to oppose each other with the fluid stream or the liquid droplets imaged by the imaging device interposed therebetween.
- the micro-particle sorting apparatus may further comprise at least one collection container configured to collect the fluid stream and capable of moving at least in a direction parallel to the imaging direction of the imaging device.
- the control unit may be configured to adjust the position of the collection container based on information regarding a deviation of the orbital direction of the fluid stream obtained by focusing on at least two parts in the image of the fluid stream.
- the focusing of the two parts may comprise a focusing of two end portions of the fluid stream in the image of the fluid stream.
- control unit may be configured to determine the diameter of the orifice based on the width of the fluid stream detected in the perpendicular direction to the trajectory direction of the fluid stream of the image imaged by the imaging device.
- micro-particle sorting apparatus may be implemented in any combination.
- Embodiments also include a method of determining a trajectory of a fluid stream or liquid droplets of a micro-particle sorting apparatus.
- the method may comprise, in order, acts of obtaining an image of an ejected fluid stream or liquid droplets, detecting a center line from contrasted points within the image, comparing the center line with preset reference line information, and displaying the image.
- the contrasted points may be located centrally within the ejected fluid stream or liquid droplets.
- the trajectory of the fluid stream S (or the liquid droplets) can be automatically determined.
- highly precise analysis can be simply performed.
Abstract
A flow cytometer includes apparatus for evaluating a trajectory of an ejected stream that carries micro-particles. The stream may be ejected from a micro-orifice of a micro-fluidic chip. The apparatus includes an imaging device and at least one processor configured to evaluate a trajectory of the ejected stream in at least two directions, e.g., a focusing direction and a direction transverse to the focusing direction. Based upon a detected trajectory, the system may execute an alarm function if the trajectory indicates an abnormal condition, or may move sample collection containers to accommodate for measured deviations in the trajectory of the ejected stream.
Description
- The present technology relates to a micro-particle sorting apparatus and a method of determining a trajectory of an ejected stream of the micro-particle sorting apparatus. In particular, the present technology relates to a micro-particle sorting apparatus which automatically determines a trajectory of a fluid stream or the like carrying micro-particles that is ejected from an orifice.
- There has been a micro-particle sorting apparatus (for example, a flow cytometer) which detects characteristics of micro-particles such as cells optically, electrically or magnetically, and fractionates or sorts the micro-particles so as to collect only micro-particles having predetermined characteristics.
- In fractionating the micro-particles with the flow cytometer, first, a fluid stream (laminar flow of sample liquid and sheath liquid containing cells) is generated from an orifice formed in a flow cell, a vibration is applied to the orifice to transform the fluid stream into a form of liquid droplets, and electric charge is applied to the liquid droplets. Then, the movement direction of the liquid droplets containing the micro-particles discharged from the orifice is electrically controlled to collect target micro-particles having the desired characteristics and non-target micro-particles having characteristics other than those desired are sorted into separate collection containers.
- For example, in Japanese Unexamined Patent Application Publication No. 2010-190680, which is incorporated herein by reference, describes a microchip flow cytometer according to one embodiment as, “a micro-particle sorting apparatus including: a microchip in which a flow path through which liquid containing micro-particles flows and an orifice through which the liquid flowing through the flow path is discharged into a space outside the chip are disposed; a vibrating element configured to transform the liquid into a form of liquid droplets and discharge the liquid droplets in the orifice; charge means for adding an electric charge to the discharged liquid droplets; optical detection means for detecting the optical characteristics of micro-particles which flow through the flow path; a pair of electrodes provided so as to be opposed to each other with the moving liquid droplets interposed therebetween along a movement direction of the liquid droplets discharged into the space outside the chip; and two or more containers that collect the liquid droplets passing through between the pair of electrodes”.
- In a micro-particle sorting apparatus, it is desirable to design fluid collection such that a fluid stream or liquid droplets generated from an orifice formed in a flow cell or a microchip enter inside a collection container. Accordingly, it is necessary to prevent a deviation of the fluid stream or the liquid droplets from an assumed direction. In the related art, the prevention of the deviation has been performed by checking an ejected fluid stream or the like with visual observation of a user, and problems regarding reliability and stability were generated and depended upon the experience level of the user. Further, performing the determination of the deviation with visual observation was extremely complicated in the configuration of the apparatus.
- It is desirable to provide a micro-particle sorting apparatus capable of automatically detecting a deviation of a trajectory of an ejected fluid stream or liquid droplets that carries the micro-particles.
- In the present technology, the term “micro-particles” broadly includes biologically-relevant micro-particles such as cells, microorganisms, liposomes, and the like, and synthetic particles such as latex particles, gel particles, industrial particles, and the like.
- The biologically-relevant micro-particles include chromosomes, liposomes, mitochondria, organelles, and the like configuring various cells. Cells include animal cells (hematopoietic cells and the like) and plant cells. Microorganisms include bacteria such as coli bacteria, viruses such as tobacco mosaic virus, and fungi such as yeast cells. Biologically-relevant micro-particles include nucleic acids, proteins, and biologically-relevant macromolecules such as a complex thereof. Industrial particles may be organic or inorganic high polymer materials, metals and the like, for example. Organic polymer materials include polystyrene, styrene-divinylbenzene, polymethylmethacrylate and the like. Inorganic polymer materials include glasses, silica, magnetic materials, and the like. Metals include gold colloids, aluminum and the like. The shape of the micro-particles is generally spherical, however it may be non-spherical and the size, weight and the like thereof are not particularly limited.
- According to the present technology, a micro-particle sorting apparatus which is capable of automatically detecting the deviation of the trajectory of an ejected fluid stream or liquid droplets is provided. According to some embodiments, a flow cytometer comprises a micro-orifice configured to eject a fluidic stream, and an imaging device configured to image an ejected stream, wherein the ejected stream is at least a portion of the fluidic stream ejected from the micro-orifice. The flow cytometer may further include at least one processor that is configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image. The ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets. The micro-orifice may be an exit orifice of a micro-fluidic chip.
- In some embodiments, the flow cytometer may further comprise electrostatic deflection apparatus configured to deflect the liquid droplets, and charging apparatus configured to apply charge to individual liquid droplets so that individual liquid droplets carrying micro-particles may be sorted according to pre-selected sorting criteria. In some embodiments, the flow cytometer may further comprise automated focusing apparatus, wherein the at least one processor is further configured to measure a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and control the focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image. The focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- According to some embodiments, the at least one processor may be configured to evaluate the trajectory of the ejected stream based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream. The at least one processor may be configured to evaluate the trajectory by identifying a line that connects the one or more contrasted spots. In some embodiments, the at least one processor may be configured to compute an angle associated with the trajectory of the ejected stream. The computed angle may be a measure of deviation of the ejected stream from a predetermined direction. According to some embodiments, the at least one processor may be further configured to detect an abnormality in operation of the flow cytometer if the computed angle is greater than a predetermined threshold value. The detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice. In some embodiments, the at least one processor may be further configured to execute an alerting function if the computed angle is greater than a predetermined threshold value.
- In some embodiments, the flow cytometer may further comprise movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- According to some embodiments, the at least one processor may be configured to calculate the trajectory of the ejected stream in a focus direction based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream. The first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream. In some embodiments, the at least one processor may be configured to calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- In some embodiments, the at least one processor may be further configured to identify a width of the ejected stream in the received image and determine a diameter of the micro-orifice based upon the identified width of the ejected stream.
- The foregoing embodiments and features of a flow cytometer may be implemented in a flow cytometer in any combination.
- Embodiments also include a trajectory evaluation system for a flow cytometer. The trajectory evaluation system may comprise an imaging device configured to image the ejected stream, wherein the ejected stream is at least at portion of a fluidic stream ejected from a micro-orifice of the flow cytometer. The trajectory evaluation system may further comprise at least one processor configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image. The ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets. The micro-orifice may be an exit orifice of a micro-fluidic chip.
- According to some embodiments, the trajectory evaluation system may include automated focusing apparatus, wherein the at least one processor is further configured to measure a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and control the focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image. The focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- According to some embodiments, the at least one processor may be configured to evaluate the trajectory of the ejected stream based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream. The at least one processor may be configured to evaluate the trajectory by identifying a line that connects the one or more contrasted spots. In some embodiments, the at least one processor may be configured to compute an angle associated with the trajectory of the ejected stream. The computed angle may be a measure of deviation of the ejected stream from a predetermined direction. According to some embodiments, the at least one processor may be further configured to detect an abnormality in operation of a flow cytometer if the computed angle is greater than a predetermined threshold value. The detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice. In some embodiments, the at least one processor may be further configured to execute an alerting function if the computed angle is greater than a predetermined threshold value.
- In some embodiments, the trajectory evaluation system may provide a signal for moving movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- According to some embodiments, the at least one processor may be configured to calculate the trajectory of the ejected stream in a focus direction based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream. The first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream. In some embodiments, the at least one processor may be configured to calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- In some embodiments, the at least one processor may be further configured to identify a width of the ejected stream in the received image and determine a diameter of the micro-orifice based upon the identified width of the ejected stream.
- The foregoing embodiments and features of a trajectory evaluation system may be implemented in a trajectory evaluation system for a flow cytometer in any combination.
- Embodiments also include a method of measuring a trajectory of an ejected stream in a flow cytometer. The method may comprise an act of imaging, with an imaging device, the ejected stream, wherein the ejected stream is at least a portion of a fluidic stream ejected from a micro-orifice of the flow cytometer. The method may further include receiving, by at least one processor, an image of the ejected stream imaged by the imaging device, and processing, by the at least one processor, the received image to detect one or more contrasted spots located centrally within the ejected stream and to evaluate a trajectory of the ejected stream. The ejected stream may comprise a continuous liquid stream or comprise a stream of separated liquid droplets. The micro-orifice may be an exit orifice of a micro-fluidic chip.
- According to some embodiments, the method of measuring a trajectory may include an act of automated focusing, wherein the at least one processor measures a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and controls focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image. The focus of the received image may be changed according to a contrast ratio computed from the first and second brightness levels.
- According to some embodiments, the method of measuring a trajectory may comprise evaluating the trajectory of the ejected stream, by the at least one processor, based upon an arrangement of one or more contrasted spots detected within the image of the ejected stream. The at least one processor may evaluate the trajectory by identifying a line that connects the one or more contrasted spots. In some embodiments, the at least one processor may compute an angle associated with the trajectory of the ejected stream. The computed angle may be a measure of deviation of the ejected stream from a predetermined direction. According to some embodiments, the at least one processor may detect an abnormality in operation of a flow cytometer if the computed angle is greater than a predetermined threshold value. The detected abnormality may be associated with or attributed to a micro-fluidic chip having the micro-orifice. In some embodiments, the at least one processor may execute an alerting function if the computed angle is greater than a predetermined threshold value.
- In some embodiments, the method of measuring a trajectory may include providing a signal, by the at least one processor, for moving movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
- According to some embodiments, the method of measuring a trajectory may include calculating the trajectory of the ejected stream in a focus direction, by the at least one processor, based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream. The first focus condition may be evaluated by focusing a first contrasted spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second contrasted spot centrally in the ejected stream near a second end of the ejected stream. In some embodiments, the at least one processor may calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
- In some embodiments, the method of measuring a trajectory may further comprise identifying, by the at least one processor, a width of the ejected stream in the received image and determining a diameter of the micro-orifice based upon the identified width of the ejected stream.
- The foregoing embodiments and features of a method of measuring a trajectory of an ejected stream in a flow cytometer may be implemented in flow cytometer in any combination.
- The foregoing and other aspects, embodiments, and features of the present teachings can be more fully understood from the following description in conjunction with the accompanying drawings.
- The skilled artisan will understand that the figures, described herein, are for illustration purposes only. It is to be understood that in some instances various aspects of the embodiments may be shown exaggerated or enlarged to facilitate an understanding of the embodiments. In the drawings, like reference characters generally refer to like features, functionally similar and/or structurally similar elements throughout the various figures. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the teachings. The drawings are not intended to limit the scope of the present teachings in any way.
-
FIG. 1 is a schematic diagram illustrating a configuration of a sorting system of a micro-particle sorting apparatus (flow cytometer) according to an embodiment of the present technology. The flow cytometer may be configured as a microchip flow cytometer. -
FIGS. 2A and 2B are schematic diagrams illustrating a configuration of an example of a microchip which is mountable on a flow cytometer. -
FIGS. 3A to 3C are schematic diagrams illustrating a configuration of an orifice of a microchip. -
FIG. 4 is a flowchart illustrating steps for determining a trajectory of a fluid stream or the like of a flow cytometer. -
FIGS. 5A and 5B are pictures showing an example of images before and after focusing which are imaged by a droplet camera of a flow cytometer. -
FIGS. 6A and 6B are schematic diagrams showing images of fluid streams having different widths from each other imaged by a droplet camera of a flow cytometer. -
FIGS. 7A and 7B are pictures showing an example of images of a fluid stream and liquid droplets imaged by a droplet camera of a flow cytometer. -
FIGS. 8A and 8B are pictures showing an example of images of a fluid stream and liquid droplets imaged by a droplet camera of a flow cytometer. -
FIGS. 9A and 9B are pictures showing an example of images of liquid droplets imaged by a droplet camera of a flow cytometer. -
FIGS. 10A and 10B are schematic diagrams showing an example of images of fluid streams imaged by a droplet camera of a flow cytometer. - The features and advantages of the present embodiments will become more apparent from the detailed description set forth below when taken in conjunction with the drawings.
- Hereinafter, an embodiment according to the present technology will be described with reference to the drawings. The embodiment, which will be described hereinafter, is an example of the representative embodiments of the present technology, and the scope of the present technology is not narrowed by the embodiment. The description will be in the following order.
- 1. Apparatus Configuration of Micro-particle Sorting Apparatus according to Embodiment of Present Technology
- 1-1. Chip Loading Module
- 1-2. Microchip
- 1-3. Deflection Plate
- 1-4. Collection Unit
- 1-5. Droplet Camera
- 1-6. Control Unit and the like
- 2. Method of Determining a Stream Trajectory of Micro-particle Sorting Apparatus according to Another Embodiment of Present Technology
- 2-1. Fluid Stream Generating Step S1
- 2-2. Droplet Camera Z Axis Scanning and Fluid Stream Imaging Step S2
- 2-3. Focusing Step S3
- 2-4. Center Line Detecting Step S4
- 2-5. Displaying Step S5
- 2-6. Orbital Direction Determining Step S6
- 2-6-1. Z Axis Direction Determining Step S61
- 2-6-2. X Axis Direction Determining Step S62
- 2-7. Alerting Step S7
- 2-8. Collection Tube Moving and Aligning Step S8
- 3. Various Additional Embodiments
-
FIG. 1 is a schematic diagram illustrating a configuration of a sorting system of a micro-particle sorting apparatus 1 (hereinafter, also referred to as a “flow cytometer 1”) according to an embodiment of the present technology. According to one embodiment, the flow cytometer is configured as a microchip flow cytometer. - 1-1. Chip Loading Module
- Reference numeral 11 in the drawing denotes a chip loading module storing a
microchip 2. The chip loading module 11 includes a chip loading unit which performs transportation to store themicrochip 2 inserted from the outside to a predetermined position, and a transporting liquid connecting unit which supplies sample liquid, sheath liquid, and the like including micro-particles to the stored microchip 2 (both not shown). In addition, the chip loading module 11 includes a chip vibrating unit which is formed in themicrochip 2, and applies a vibration to theorifice 21 generating laminar flow (flow stream S) of sample liquid and sheath liquid to transform the fluid stream S into a form of liquid droplets and discharge the liquid droplets, and an electric charge unit which applies an electric charge to the discharged liquid droplets (both not shown). - 1-2. Microchip
-
FIGS. 2A to 3C show an example of themicrochip 2 which is mountable on theflow cytometer 1.FIG. 2A shows a schematic diagram of the upper surface andFIG. 2B shows a cross-sectional schematic diagram taken along the line IIB-IIB ofFIG. 2A .FIGS. 3A to 3C are diagrams schematically illustrating one configuration of theorifice 21 of themicrochip 2.FIG. 3A shows a schematic diagram of the upper surface,FIG. 3B shows a cross-sectional schematic diagram, andFIG. 3C shows a plan diagram.FIG. 3B is a cross-sectional diagram taken along the line IIIB-IIIB ofFIG. 2A . - The
microchip 2 may be formed by bondingsubstrate layers sample flow path 22 is formed. Thesample flow path 22 on the substrate layers 2 a and 2 b can be formed by injection molding of thermoplastic resin using mold. For the thermoplastic resin, existing plastics of the related art as materials of the microchip such as polycarbonate, polymethylmethacrylate resin (PMMA), cyclic polyolefins, polyethylene, polystyrene, polypropylene, polydimethylsiloxane (PDMS) or the like may be used. - The sample liquid is introduced to a
sample inlet 23 from the transporting liquid connecting unit and joins with the sheath liquid which is introduced to asheath inlet 24 from the transporting liquid connecting unit to transport the liquid to thesample flow path 22. The sheath liquid introduced from thesheath inlet 24 is transported by dividing into two directions. Then, in the joining portion with the sample liquid introduced from thesample inlet 23, the sheath liquid joins with the sample liquid so as to interpose the sample liquid from two directions. Accordingly, in the joining portion, three-dimensional laminar flow is formed in which the laminar flow of the sample liquid is positioned at the center of the laminar flow of the sheath liquid. -
Reference numeral 25 denotes a suction flow path for removing clogging and bubbles by causing a negative pressure inside thesample flow path 22 to counterflow temporarily, when clogging and bubbles are generated in thesample flow path 22. At the one end of thesuction flow path 25, asuction outlet 251 which is connected to a negative pressure source such as a vacuum pump or the like through the transporting liquid connecting unit is formed, and another end thereof is connected to thesample flow path 22 in a communicatingport 252. - The laminar flow width of the three-dimensional laminar flow may be formed to be narrowed down in narrowing units 261 (see
FIGS. 2A and 2B ) and 262 (seeFIGS. 3A to 3C) so that the area of the perpendicular cross section with respect to the transporting liquid direction becomes small gradually or in steps from the upstream to the downstream in the transporting liquid direction. After that, the three-dimensional laminar flow becomes the fluid stream S (seeFIG. 1 ) and is discharged from theorifice 21 provided at one end of the flow path. InFIG. 1 , the discharging direction of the fluid stream S from theorifice 21 is shown as the positive Y axis direction. - The characteristics of the micro-particles may be detected between the narrowing
unit 261 and thenarrowing unit 262 of thesample flow path 22. For example, in optical detection by a light irradiation detecting unit (now shown), a laser is emitted with respect to the micro-particles which are arranged in a line in the center of the three-dimensional laminar flow to flow inside thesample flow path 22, and scattering light and fluorescence generated from the micro-particles are detected by one or more light detectors. - A connecting unit of the
sample flow path 22 and theorifice 21 is set as astraight unit 27 formed to be linear. Thestraight unit 27 functions for ejecting the fluid stream S from theorifice 21 linearly in the positive Y axis direction. - The fluid stream S ejected from the
orifice 21 may be transformed into a form of liquid droplets by the vibration applied to theorifice 21 by a chip vibrating unit. Theorifice 21 is opened in the end surface direction of the substrate layers 2 a and 2 b, and a cut-outportion 211 is provided between the opening position and the end surface of the substrate layers. The cut-outportion 211 is formed by cutting out the substrate layers 2 a and 2 b between the opening position of theorifice 21 and the end surface of the substrates so that a diameter L of the cut-out portion 221 is larger than adiameter 1 of the opening of the orifice 21 (seeFIG. 3C ). It is desirable that the diameter L of the cut-outportion 211 be formed to be larger by more than double thediameter 1 of the opening of theorifice 21 so as not to interrupt the movement of the liquid droplets discharged from theorifice 21. - 1-3. Deflection Plates
-
Reference numerals FIG. 1 denote a pair of deflection plates which are arranged to oppose each other by interposing the fluid stream S (or the discharged liquid droplets) which is ejected from theorifice 21 and imaged by adroplet camera 4 which will be described later. Thedeflection plates orifice 21 by an electric force interacting with electric charge applied to the liquid droplets. In addition, thedeflection plates orifice 21 by an electric force interacting with electric charge applied to the fluid stream S. InFIG. 1 , the opposing direction of thedeflection plates - 1-4. Collection Unit
- In the
flow cytometer 1, the fluid stream S (or liquid droplets D thereof) may be collected in any of a plurality of collection tubes (collection containers) 3 which are arranged in a line in the opposing direction (X axis direction) of thedeflection plates 12 and 12 (seeFIG. 1 ). Thecollection tubes 3 may be general-purpose plastic tubes or experimental glass tubes. The number of thecollection tubes 3 is not particularly limited, but the embodiment shows a case of arranging five collection tubes. The fluid stream S generated from theorifice 21 is introduced to any one of the fivecollection tubes 3 depending on the existence or non-existence, or the size of the electric force acting between thedeflection plates - The
collection tubes 3 may be disposed in acollection tube container 31 in an exchangeable manner. Thecollection tubes 3 are disposed in the movement direction (X axis direction) shown as an arrow F1 inFIG. 1 in a movable manner. For example, thecollection tubes 3 may be disposed so that only thecollection tubes 3 move in the X axis direction in a state where thecollection tube container 31 is fixed, or thecollection tubes 3 may be disposed movably with the movement of thecollection tube container 31. - The
collection tube container 31 may be disposed on aZ axis stage 32 which is configured to be movable in a direction (Z axis direction) perpendicular to the discharging direction (Y axis direction) of the fluid stream S from theorifice 21 and the opposing direction (X axis direction) of thedeflection plates FIG. 1 denotes the movement direction of theZ axis stage 32.Reference numeral 321 in the drawing denotes a waste liquid port provided on theZ axis stage 32. In theflow cytometer 1, thecollection tube container 31 and theZ axis stage 32 configure acollection unit 33 which is driven by a Z axis motor (not shown). - 1-5. Droplet Camera
- A
droplet camera 4 may be any suitable camera (CCD camera, CMOS image sensor or the like) for imaging the fluid stream S ejected from theorifice 21 of themicrochip 2 or the liquid droplets discharged therefrom (seeFIG. 1 ). Thedroplet camera 4 may be designed to be able to perform automated focusing under the control of at least one processor on the captured image of the fluid stream S or the liquid droplets. The image captured by thedroplet camera 4 may be displayed on the display unit such as a display, and used for a user to check for the formation state (size, shape, intervals and the like of the liquid droplets) of the liquid droplets of theorifice 21. - In the
flow cytometer 1, the trajectories of the fluid stream S (or liquid droplets) ejected from theorifice 21 are different depending on the individual differences of the mountedmicrochips 2, and the position of the fluid stream S can be changed in the Z axis direction (and X axis direction) in the drawing, at each time of exchanging themicrochip 2. Continuing ejecting the fluid stream S or continuing discharging the liquid droplets may result in the degradation or the like of themicrochip 2, so that the position of the fluid stream S (or the liquid droplets) can be changed over time in the Z axis direction (and X axis direction) in the drawing. Thedroplet camera 4 also functions for detecting such position change of the fluid stream S (or the liquid droplets) in the Z axis direction (and X axis direction). - 1-6. Control Unit and the Like
- In addition to the above described configuration, the
flow cytometer 1 includes a light irradiation detecting unit for detecting the optical characteristics of micro-particles, a data analysis unit for determining the characteristics, a tank unit which stores the sample liquid and the sheath liquid, and a control unit for controlling each configuration thereof, which are included in general flow cytometers. - The control unit may be configured by a general-purpose computer including at least one CPU, a memory or a hard disk and the like, and an OS. Machine-readable instructions that may be executed by the at least one CPU may be stored in memory and, when executed by the at least one CPU, specially adapt the computer for executing each step of the position control, which will be described later, and other processes of the flow cytometer.
- The light irradiation detecting unit may be configured by a laser light source, an irradiation system which includes a condensing lens, a dichroic minor, a bandpass filter and the like which condense and emit the laser with respect to the micro-particles, and a detecting system which detects the measuring target light generated from the micro-particles by excitation of the laser. The detecting system may be configured by an area imaging device or the like such as a PMT (photomultiplier tube), a CCD, or a CMOS element.
- The measuring target light which is detected by the detecting system of the light irradiation detecting unit is the light which is generated from the micro-particles by the emission of the measuring light, and can be scattered light such as forward-scattered light, backward-scattered light, Rayleigh-scattered or Mie scattered light, or fluorescence. The above measuring target light is converted into electrical signals, output to the control unit and provided for determining the optical characteristics of the micro-particles.
- The
flow cytometer 1 may magnetically or electrically detect the characteristics of the micro-particles. In this case, microelectrodes are arranged to oppose each other in thesample flow path 22 of themicrochip 2, and a resistance value, a capacitance value, an inductance value, impedance, a changing value of the electric field between the electrodes, or the change in magnetization, magnetic field, and the like are measured. - 2-1. Fluid Stream Generating Step S1
-
FIG. 4 is a flowchart illustrating steps for determining the trajectory of the fluid stream S (or the liquid droplets) of theflow cytometer 1, according to one embodiment. The steps for determining the trajectory include processes of a “fluid stream generating step S1,” a “droplet camera Z axis scanning and fluid stream imaging step S2,” a “focusing step S3,” a “center line detecting step S4,” a “displaying step S5,” an “orbital direction determining step S6,” an “alerting step S7,” and a “collection tube moving and aligning step S8.” Hereinafter, each process will be described. - First, in the fluid stream generating step S1, the transporting liquid connecting unit starts transporting the sample liquid and the sheath liquid to the
sample inlet 23 and thesheath inlet 24 of themicrochip 2, and a fluid stream S is ejected from the orifice 21 (seeFIG. 4 ). The control unit outputs the signals to the transporting liquid connecting unit and starts transporting the sample liquid and the sheath liquid. The fluid stream S ejected from theorifice 21 may be collected in thewaste liquid port 321 and disposed of. - In this step S1, the chip vibrating unit applies the vibration to the
orifice 21, and the liquid droplets may be discharged instead of a continuous fluid stream S from the orifice, so that the liquid droplets can be collected in thewaste liquid port 321 and disposed of. - 2-2. Droplet Camera Z Axis Scanning and Fluid Stream Imaging Step S2
- In the step S2, the control unit outputs the signals to the
droplet camera 4 and thedroplet camera 4 which receives the signals may be moved in the Z axis direction (seeFIG. 4 ), for example, to center an image of the stream. Then, the control unit outputs the signals to thedroplet camera 4, and thedroplet camera 4 which receives the signals performs imaging of the fluid stream S (or the liquid droplets). - 2-3. Focusing Step S3
- In the step S3, in a case where the image of the fluid stream S (or the liquid droplets) is detected, by the control unit, the focusing may be performed in the X axis direction when imaging the image of the fluid stream S (or the liquid droplets) by the droplet camera 4 (see
FIG. 4 ). The image of the fluid stream S (or the liquid droplets) imaged by thedroplet camera 4 may be output to the control unit, and the control unit may perform focusing control until detecting the contrasted or bright points in the image in the focusing step S3. Herein, the bright points denote one or a plurality of pixels having higher brightness than a predetermined threshold value in the image of the fluid stream S (or the liquid droplets) imaged by thedroplet camera 4. A contrasted point or spot may be a spot having a luminance or color significantly different (e.g., greater than about 10% variation) from a background luminance or color around the spot. For example, a contrasted spot may be a gray spot on a white background, a yellow spot on a red background, a white spot on a black background, etc. in a recorded image. -
FIG. 5A represents a picture showing an example of a state before the focusing of the imaged liquid droplets is performed (seeFIG. 5A ), andFIG. 5B represents a picture showing an example of a state after the focusing of the liquid droplets is performed (seeFIG. 5B ). As shown inFIG. 5B , since the focusing of the image P is performed, it is possible to detect at least one bright point B in the center position of each liquid droplet D. Even in a case where the fluid stream S is ejected instead of the liquid droplets D from the orifice, it is possible to detect at least one bright point B in the center portion along the trajectory of the fluid stream S in the same manner. Accordingly, in the step S4, the focusing of thedroplet camera 4 is executed until at least one bright point B is detected in the captured image P. At that time, in a case where the contrast ratio of the image P is in a predetermined range, the control unit can determine whether the image P is in a focused state. - When the fluid stream S is imaged by the
droplet camera 4, the control unit may determine the diameter of the orifice, based on the width of the fluid stream S detected in the direction perpendicular (Z axis direction) to the trajectory of the fluid stream S in the captured image P. -
FIGS. 6A and 6B show schematic diagrams of two captured images which have a different width of the fluid stream S from each other (FIGS. 6A and 6B ). The control unit may be configured to determine accurately that the diameter of the orifice is 100 μm or the like, for example, by evaluating the width of the fluid stream S shown inFIG. 6A based on information stored in a memory unit. In the example shown inFIG. 6B , which has different width of the fluid stream S from that shown inFIG. 6A , the control unit may be configured to determine accurately that the diameter of the orifice is 70 μm or the like, for example, by evaluating the width of the fluid stream S based on the information stored in the memory unit. The control unit may record or display the determined diameter of the orifice as the diameter of the orifice of the chip used in theflow cytometer 1. Accordingly, a manual setting or recording of the diameter of the orifice by a user is not necessary, and thus it is possible to prevent setting mistakes such as mis-setting or mis-recording the diameter of the orifice. - 2-4. Center Line Detecting Step S4
- In the step S4, the control unit may detect a center line of the fluid stream S from one or more bright points in the image of the fluid stream S (or the liquid droplets D) imaged by the
droplet camera 4, and may compare preset center line information with the detected center line (seeFIG. 4 ). -
FIGS. 7A and 7B show states when a center line L of the fluid stream S (or the liquid droplets D) is detected in the captured image. When the fluid stream S is ejected from the orifice, the control unit may be configured to detect the straight line formed by the plurality of bright points displayed along the ejecting direction of the fluid stream S in the image of the fluid stream S imaged by thedroplet camera 4 as the center line L. In detail, as shown inFIG. 7A , the control unit may identify the bright points B in the captured image P of the fluid stream S as the center line L. - When the liquid droplets D are discharged from the orifice, the control unit may be configured to detect a straight line formed by connecting one or more bright points displayed in each of the liquid droplets D as the center line L. In detail, as shown in
FIG. 7B , when the liquid droplets D are imaged, the control unit may identify a line formed by connecting the bright points of each of the liquid droplets as the center line L of the liquid droplets. In this case, according to the connecting method of the bright points of each of the liquid droplets D, when a plurality of center lines L can be generated, the control unit may set a line which most closely approximates the center line information which will be described later, as the center line. - 2-5. Displaying Step S5
- In the step S5, the control unit can display the captured image on a display unit such as a display monitor (see
FIG. 4 ). - As shown in
FIGS. 7A and 7B , the control unit can arrange and display the fluid stream S (seeFIG. 7A ) or the liquid droplets D (seeFIG. 7B ) of the captured image in the center of such a display based on the center line L which is described above. In more detail, for example, first, the control unit may align thedroplet camera 4 in the Z axis direction. The control unit may perform alignment based on the captured image P, until the number of the pixels of the positive direction side and the negative direction side of the Z axis direction become the same by setting the center line L as the boundary between positive side and negative side pixels. - Accordingly, in the
flow cytometer 1, the image P of the fluid stream S (or the liquid droplets D) can be automatically aligned and displayed in the center of the display. - 2-6. Orbital Direction Determining Step S6
- In the step S6, the control unit may determine the trajectory of the fluid stream S (or the liquid droplets D) (see
FIG. 4 ). In more detail, the control unit may determines a deviation of the trajectory in the Z axis direction and also a deviation of the trajectory in the X axis direction. Hereinafter, the processes of “Z axis direction determining step S61” and “X axis direction determining step S62” are included. Each process will be described later. - 2-6-1. Z Axis Direction Determining Step S61
- In the step S61, the control unit may determine a trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction.
- As depicted in
FIGS. 7A and 7B , the control unit may compare the center line L and predetermined center line information stored in the memory unit in advance. With respect to the fluid stream S (or the liquid droplets D), the center line L is detected as described above. The predetermined center line information may be information representing a straight line perpendicular to XZ plane stored in the memory unit in advance, and may further represent a line which makes the number of pixels of the positive direction side and the negative direction side of the Z axis direction the same by setting the predetermined center line as the boundary, in the captured image. - Herein, the comparison between the center line L and the predetermined center line information I stored in the memory unit in advance will be described while further referring to
FIGS. 8A and 8B , in addition toFIGS. 7A and 7B .FIGS. 8A and 8B also show states where the determined center line L of the fluid stream S (or the liquid droplets D) is detected in the captured image in the same manner asFIGS. 7A and 7B . - In the example shown in
FIG. 8A , the center line L is deviated by θ1 degrees in the YZ plane with respect to the predetermined center line information I (seeFIG. 8A ). In the same manner, in the example shown inFIG. 8B , the center line is deviated by θ2 degrees in the YZ plane when compared to the predetermine center line information I (seeFIG. 8B ). In the example shown inFIG. 8A , the process of the display step S5, for making the number of the pixels of the positive direction side and the negative direction side of the Z axis direction the same by setting the center line L as the boundary by the control unit is omitted. - Meanwhile, in a case of the example shown in
FIGS. 7A and 7B , the control unit may determine that there is nearly no deviation of the center line L in the YZ plane with respect to the predetermined center line information I (seeFIGS. 7A and 7B ). - The control unit may also be configured to determine that an inclination angle (e.g., the angles θ1, or θ2) with respect to the center line information I of the center line L detected based on the comparison of the center line L and the center line information I, exceeds a predetermined threshold value, and determine that the microchip is abnormal. As described above, the control unit can determine the deviation of the trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction by comparing the center line information I and the center line L, and when the trajectory is deviated, the control unit can automatically determine that the microchip or the like is in a malfunction state (abnormal state of clogging or the like). An inclination angle that may result in an abnormal determination may be an inclination angle greater than 0.5 degree in some embodiments, greater than 1 degree in some embodiments, greater than 2 degrees in some embodiments, greater than 5 degrees in some embodiments, greater than 10 degrees in some embodiments, or greater than 20 degrees in some embodiments. An abnormal inclination angle may be an angle at which the ejected stream will no longer be captured by a collection vessel.
- 2-6-2. X Axis Direction Determining Step S62
- In the step S62, the control unit may determine a trajectory of the fluid stream S (or the liquid droplets D) in the X axis direction (see
FIG. 4 ). -
FIGS. 9A and 9B show pictures of an example of the images of the liquid droplets in which the focusing may be performed. As shown inFIG. 9A , when the trajectory of the liquid droplets D is not deviated in the X axis direction, since the focusing of thedroplet camera 4 is performed based on the signals of the control unit, a focused region R1 is detected for the length of the stream in the image, while a non-focused region R2 is not detected. - Meanwhile, as shown in
FIG. 9B , when the trajectory of the liquid droplets D is deviated in the X axis direction, since the focusing of thedroplet camera 4 is performed based on the signals of the control unit, a focused region R1 is detected for a portion of the stream length and a non-focused region R2 is also detected. - Since the control unit detects both the non-focused region R2 and the focused region R1 in the image P, the abnormity of the microchip or the like can be determined. Accordingly, when identifying the existence of the non-focused region R2 in the image of the fluid stream S or the liquid droplets D and confirming the existence of the non-focused region R2, the control unit determines that the trajectory of the fluid stream S or the liquid droplets D is deviated in the X axis direction. Therefore, in the
flow cytometer 1, when the trajectory is deviated in the X axis direction, the control unit may automatically determine that the microchip or the like is in a malfunction state (abnormity state of clogging or the like). -
FIGS. 10A and 10B show schematic diagrams of the image of fluid stream S in which at least one bright point is detected. As shown inFIGS. 10A and 10B , for example, the control unit may perform focusing on a negative direction side of the Y axis direction, and at least one bright point B may be detected (seeFIG. 10A ). Further, the control unit may perform focusing on a positive direction side of the Y axis direction, and at least one bright point B may be detected at a second focus position different from a first focus position found for the at least one bright point B shown inFIG. 10A (seeFIG. 10B ). Accordingly, the control unit can perform the focusing on two portions of the end portions (end portion of the positive Y axis direction side and the end portion of the negative Y direction side) of the fluid stream S in the image P of the fluid stream S. Thus, the control unit can obtain position information corresponding to the deviation of the trajectory in the X axis direction, and in the collection tube moving and aligning step S8 which will be described later, the aligning of thecollection tubes 3 in the X axis direction can be performed by using the detected position information. - 2-7. Alerting Step S7
- In the step S7, after evaluating stream or droplet trajectories in the Z axis direction and/or the X axis direction, the control unit may determine that the inclination or deviation angle exceeds a predetermined threshold value. In response, the control unit may perform alerting with respect to a user (see
FIG. 4 ). In this case, various methods such as, a method for displaying a light or a message by a display unit such as a display, or a method for providing an output unit in theflow cytometer 1 and alerting by an audio output or the like, can be used as a method for alerting a user. Thus, the user can check for the, malfunction, breakage, or the like of the chip. - 2-8. Collection Tube Moving and Aligning Step S8
- In the collection tube moving and aligning step S8, the control unit may perform positioning of the
collection tubes 3 based on the position information corresponding to the deviation of the trajectory in the X axis direction described above (seeFIG. 4 ). In detail, the information regarding the trajectory of the fluid stream S (or the liquid droplets D) in the X axis direction is converted into the position information of thecollection tubes 3 in the same direction, and thecollection tubes 3 are moved to the position corresponding to the converted position information. Accordingly, thecollection tubes 3 disposed in thecollection tube container 31 and the fluid stream S are aligned in the X axis direction, and it is possible for the ejected fluid stream S to reach thecollection tubes 3 precisely. - In addition, in the collection tube moving and aligning step S8, the control unit may perform positioning of the
collection tube container 31 based on the position information obtained by the aligning of the Z axis direction described above. In detail, the information regarding the trajectory of the fluid stream S (or the liquid droplets D) in the Z axis direction is converted into the position information of thecollection tubes 3 in the same direction, and theZ axis stage 32 is moved to the position corresponding to the converted position information. Thus, thecollection tubes 3 disposed in thecollection tube container 31 and the fluid stream S are aligned in the Z axis direction, and it is possible for the ejected fluid stream S to reach thecollection tubes 3 precisely. In the above descriptions, each process of the steps S1 to S8 has been described in order, however, the present technology is not limited to be executed in this order. For example, the process of step S7 may be executed after the process of the step S8. In some embodiments, not all steps may be implemented. In some embodiments, one or more steps may be repeated. - Additional embodiments of apparatus and related methods are also contemplated. In some embodiments, a micro-particle sorting apparatus comprises an imaging device that images a fluid stream ejected from an orifice, or liquid droplets discharged from the orifice, and a control unit. The control unit may be configured to detects a center line of the fluid stream or the plurality of the liquid droplets from contrasted points in an image of the fluid stream or the liquid droplets imaged by the imaging device, and compare the center line with preset center line information. The micro-particle sorting apparatus may further include a display unit that displays the image. According to some embodiments, the imaging device may be configured to focus the captured image, and the control unit may performs focusing on at least a part of regions of the image. The control unit may be configured to determine, based upon a contrast ratio of selected portion of the image falling in a predetermined range, that the image is in a focused state or a non-focused state. According to some embodiments, the micro-particle sorting apparatus may comprise a microchip flow cytometer in which the orifice is provided in a microchip.
- In some embodiments, the control unit may be configured to identify or set a straight line corresponding to a plurality of the contrasted points of the fluid stream displayed along the ejection direction in the image of the fluid stream imaged by the imaging device. The straight line may be identified as the center line and trajectory of the ejected fluid stream from the orifice. In some embodiments, the control unit may be configured to identify or set the straight line corresponding to a plurality of the contrasted points of the liquid droplets displayed along the ejection direction discharged from the orifice in the image of the fluid stream imaged by the imaging device, and to identify the straight line as a center line and trajectory of the ejected droplets. The control unit may be configured to determine an abnormity of the ejected fluid stream or liquid droplets by calculating an inclination value between the identified center line and a predetermined reference line. The abnormality may be determined when a comparison between the center line and the reference line exceeds a predetermined threshold value.
- In some embodiments of the micro-particle sorting apparatus, the control unit may be configured to determine an existence of a non-focused region in the image of the fluid stream or the plurality of the liquid droplets. The control unit may determine an abnormity in the image when the non-focused region and a focused region are detected in the image of the fluid stream or the plurality of the liquid droplets.
- According to some embodiments, the micro-particle sorting apparatus may comprise a pair of deflection plates that are disposed to oppose each other with the fluid stream or the liquid droplets imaged by the imaging device interposed therebetween. The micro-particle sorting apparatus may further comprise at least one collection container configured to collect the fluid stream and capable of moving at least in a direction parallel to the imaging direction of the imaging device. The control unit may be configured to adjust the position of the collection container based on information regarding a deviation of the orbital direction of the fluid stream obtained by focusing on at least two parts in the image of the fluid stream. The focusing of the two parts may comprise a focusing of two end portions of the fluid stream in the image of the fluid stream.
- In some embodiments, the control unit may be configured to determine the diameter of the orifice based on the width of the fluid stream detected in the perpendicular direction to the trajectory direction of the fluid stream of the image imaged by the imaging device.
- The foregoing embodiments and features of a micro-particle sorting apparatus may be implemented in any combination.
- Embodiments also include a method of determining a trajectory of a fluid stream or liquid droplets of a micro-particle sorting apparatus. The method may comprise, in order, acts of obtaining an image of an ejected fluid stream or liquid droplets, detecting a center line from contrasted points within the image, comparing the center line with preset reference line information, and displaying the image. The contrasted points may be located centrally within the ejected fluid stream or liquid droplets.
- As described above, in the
flow cytometer 1, the trajectory of the fluid stream S (or the liquid droplets) can be automatically determined. Thus, in theflow cytometer 1, highly precise analysis can be simply performed. - The present technology contains subject matter related to that disclosed in Japanese Priority Patent Application JP 2012-080366 filed in the Japan Patent Office on Mar. 30, 2012, the entire contents of which are hereby incorporated by reference.
- It should be understood by those skilled in the art that various modifications, combinations, sub-combinations and alterations may occur depending on design requirements and other factors insofar as they are within the scope of the appended claims or the equivalents thereof.
Claims (20)
1. A flow cytometer comprising:
a micro-orifice configured to eject a fluidic stream;
an imaging device configured to image an ejected stream, the ejected stream being at least a portion of the fluidic stream ejected from the micro-orifice; and
at least one processor configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image.
2. The flow cytometer of claim 1 , wherein the ejected stream comprises a continuous liquid stream or comprises a stream of separated liquid droplets.
3. The flow cytometer of claim 2 , further comprising:
electrostatic deflection apparatus configured to deflect the liquid droplets; and
charging apparatus configured to apply charges to individual liquid droplets so that individual liquid droplets carrying micro-particles may be sorted according to pre-selected sorting criteria.
4. The flow cytometer of claim 2 , further comprising automated focusing apparatus, wherein the at least one processor is further configured to measure a first brightness level within at least one central region of the liquid stream or liquid droplets and a second brightness level in at least one remaining portion of the liquid stream or liquid droplets and control the focusing apparatus based upon the measured first and second brightness levels so as to change a focus of the received image.
5. The flow cytometer of claim 4 , wherein the focus of the received image is changed according to a contrast ratio computed from the first and second brightness levels.
6. The flow cytometer of claim 1 , wherein the at least one processor is configured to evaluate the trajectory of the ejected stream based upon an arrangement of one or more bright spots detected within the image of the ejected stream.
7. The flow cytometer of claim 6 , wherein the at least one processor is configured to evaluate the trajectory by identifying a line that connects the one or more bright spots.
8. The flow cytometer of claim 1 , wherein the at least one processor is further configured to compute an angle associated with the trajectory of the ejected stream.
9. The flow cytometer of claim 8 , wherein the computed angle is a measure of deviation of the ejected stream from a predetermined direction.
10. The flow cytometer of claim 8 , wherein the at least one processor is further configured to detect an abnormality in operation of the flow cytometer if the computed angle is greater than a predetermined threshold value.
11. The flow cytometer of claim 10 , wherein the detected abnormality is associated with a micro-fluidic chip having the micro-orifice.
12. The flow cytometer of claim 10 , wherein the at least one processor is further configured to execute an alerting function if the computed angle is greater than a predetermined threshold value.
13. The flow cytometer of claim 1 , further comprising movable sample collection tubes that are configured to be moved in an automated manner responsive to the at least one processor determining that the trajectory of the ejected stream deviates from a predetermined trajectory.
14. The flow cytometer of claim 1 , wherein the at least one processor is configured to calculate the trajectory of the ejected stream in a focus direction based upon a first focus condition of a first portion of the ejected stream and a second focus condition of a second portion of the ejected stream.
15. The flow cytometer of claim 14 , wherein the first focus condition is evaluated by focusing a first bright spot centrally in the ejected stream near a first end of the ejected stream and the second focus condition is evaluated by focusing a second bright spot centrally in the ejected stream near a second end of the ejected stream.
16. The flow cytometer of claim 14 , wherein the at least one processor is configured to calculate the trajectory of the ejected stream based upon a difference in positions associated with the first focus condition and second focus condition.
17. The flow cytometer of claim 1 , wherein the at least one processor is further configured to identify a width of the ejected stream in the received image and determine a diameter of the micro-orifice based upon the identified width of the ejected stream.
18. The flow cytometer of claim 1 , wherein the micro-orifice is an exit orifice of a micro-fluidic chip.
19. A trajectory evaluation system for an ejected stream of a flow cytometer, the trajectory evaluation system comprising:
an imaging device configured to image the ejected stream, wherein the ejected stream is at least at portion of a fluidic stream ejected from a micro-orifice of the flow cytometer; and
at least one processor configured to receive and process an image of the ejected stream imaged by the imaging device, detect one or more contrasted spots located centrally within the ejected stream, and evaluate a trajectory of the ejected stream from the received image based upon the one or more contrasted spots.
20. A method of measuring a trajectory of an ejected stream in a flow cytometer, the method comprising:
imaging, with an imaging device, the ejected stream, wherein the ejected stream is at least a portion of a fluidic stream ejected from a micro-orifice of the flow cytometer;
receiving, by at least one processor, an image of the ejected stream imaged by the imaging device; and
processing, by the at least one processor, the received image to detect one or more contrasted spots located centrally within the ejected stream and to evaluate a trajectory of the ejected stream based upon the one or more contrasted spots.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US16/517,281 US11193874B2 (en) | 2012-03-30 | 2019-07-19 | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2012080366A JP5924077B2 (en) | 2012-03-30 | 2012-03-30 | Fine particle sorting device and method for determining orbit direction in fine particle sorting device |
JP2012080366 | 2012-03-30 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/517,281 Continuation US11193874B2 (en) | 2012-03-30 | 2019-07-19 | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
Publications (1)
Publication Number | Publication Date |
---|---|
US20130258075A1 true US20130258075A1 (en) | 2013-10-03 |
Family
ID=49234449
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/788,165 Abandoned US20130258075A1 (en) | 2012-03-30 | 2013-03-07 | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
US16/517,281 Active 2033-05-25 US11193874B2 (en) | 2012-03-30 | 2019-07-19 | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/517,281 Active 2033-05-25 US11193874B2 (en) | 2012-03-30 | 2019-07-19 | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
Country Status (3)
Country | Link |
---|---|
US (2) | US20130258075A1 (en) |
JP (1) | JP5924077B2 (en) |
CN (1) | CN103364325B (en) |
Cited By (29)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20120076349A1 (en) * | 2009-06-03 | 2012-03-29 | Hitachi High-Technologies Corporation | Flow type particle image analysis method and device |
US20140176704A1 (en) * | 2011-02-04 | 2014-06-26 | Cytonome/St, Llc | Fluid stream imaging apparatus |
US20140306122A1 (en) * | 2013-04-12 | 2014-10-16 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US9029724B2 (en) | 2012-03-30 | 2015-05-12 | Sony Corporation | Microparticle sorting device and method for controlling position in microparticle sorting device |
US9087371B2 (en) | 2012-03-30 | 2015-07-21 | Sony Corporation | Microparticle sorting device and method of optimizing fluid stream therein |
US9339823B2 (en) | 2012-03-30 | 2016-05-17 | Sony Corporation | Microparticle sorting apparatus and delay time determination method |
CN105659069A (en) * | 2013-10-16 | 2016-06-08 | 索尼公司 | Particle fractionation device, particle fractionation method, and program |
CN105980831A (en) * | 2014-02-13 | 2016-09-28 | 索尼公司 | Particle sorting apparatus, particle sorting method, program, and particle sorting system |
US20170010203A1 (en) * | 2014-02-14 | 2017-01-12 | Sony Corporation | Particle sorting apparatus, particle sorting method, and non-transitory computer-readable storage medium storing program |
US20170074775A1 (en) * | 2014-05-22 | 2017-03-16 | Sony Corporation | Particle analyzer |
US9669403B2 (en) | 2012-07-25 | 2017-06-06 | Sony Corporation | Microparticle measurement device and liquid delivery method in microparticle measurement device |
US9784659B2 (en) | 2012-11-08 | 2017-10-10 | Sony Corporation | Microparticle fractionating apparatus and method of fractionating microparticle |
US9784660B2 (en) | 2013-01-28 | 2017-10-10 | Sony Corporation | Microparticle sorting device, and method and program for sorting microparticles |
WO2017222461A1 (en) * | 2016-06-21 | 2017-12-28 | Giatrellis Sarantis | System and method for precision deposition of liquid droplets |
US9857286B2 (en) | 2013-10-17 | 2018-01-02 | Sony Corporation | Particle fractionation apparatus, particle fractionation method and particle fractionation program |
US9915599B2 (en) | 2013-02-08 | 2018-03-13 | Sony Corporation | Microparticle analysis apparatus and microparticle analysis system |
US9915935B2 (en) | 2012-03-30 | 2018-03-13 | Sony Corporation | Microchip-type optical measuring apparatus and optical position adjusting method thereof |
US9964968B2 (en) | 2013-03-14 | 2018-05-08 | Cytonome/St, Llc | Operatorless particle processing systems and methods |
US10156510B2 (en) | 2014-08-28 | 2018-12-18 | Sysmex Corporation | Particle imaging apparatus and particle imaging method |
WO2019008044A1 (en) * | 2017-07-04 | 2019-01-10 | Cytena Gmbh | Method for dispensing a liquid |
US10386287B2 (en) | 2014-09-05 | 2019-08-20 | Sony Corporation | Droplet sorting device, droplet sorting method and program |
EP3511691A4 (en) * | 2016-09-12 | 2019-12-18 | Sony Corporation | Microparticle measurement device and microparticle measurement method |
WO2020011773A1 (en) * | 2018-07-09 | 2020-01-16 | Cytena Gmbh | Method for examining a liquid sample and a dispensing apparatus |
US10605714B2 (en) | 2015-10-19 | 2020-03-31 | Sony Corporation | Image processing device, fine particle sorting device, and image processing method |
US10723497B2 (en) * | 2014-11-03 | 2020-07-28 | Vanrx Pharmasystems Inc. | Apparatus and method for monitoring and controlling the filling of a container with a pharmaceutical fluid in an aseptic environment |
US20210270721A1 (en) * | 2018-10-01 | 2021-09-02 | Hewlett-Packard Development Company, L.P. | Particle sorting using microfluidic ejectors |
US11193874B2 (en) | 2012-03-30 | 2021-12-07 | Sony Corporation | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
WO2021262359A1 (en) * | 2020-06-24 | 2021-12-30 | Becton, Dickinson And Company | Flow cytometric droplet dispensing systems and methods for using the same |
CN115414972A (en) * | 2022-08-08 | 2022-12-02 | 广东省科学院生物与医学工程研究所 | Portable coaxial focusing micro-droplet generation device and micro-droplet preparation method |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
TWI579150B (en) * | 2014-01-27 | 2017-04-21 | 國立交通大學 | Cleaning apparatus for inkjet print head |
CN107003225B (en) * | 2014-12-04 | 2020-08-18 | 贝克顿·迪金森公司 | Flow cytometry cell sorting system and method of use thereof |
US11009338B2 (en) * | 2017-04-04 | 2021-05-18 | Panasonic Intellectual Property Management Co., Ltd. | Liquid droplet measurement method and liquid droplet measurement device, and method and apparatus for manufacturing device |
WO2020091866A1 (en) * | 2018-10-30 | 2020-05-07 | Becton, Dickinson And Company | Particle sorting module with alignment window, systems and methods of use thereof |
JP7353623B2 (en) * | 2019-08-05 | 2023-10-02 | アライドフロー株式会社 | Particle separation device and particle separation method |
CN111521549B (en) * | 2020-05-13 | 2021-01-01 | 洹仪科技(上海)有限公司 | Particle sorting device and method |
US11858008B2 (en) * | 2021-03-26 | 2024-01-02 | Cytonome/St, Llc | Systems and methods for particle sorting with automated adjustment of operational parameters |
US11890872B2 (en) * | 2021-08-17 | 2024-02-06 | Wisconsin Alumni Research Foundation | Aerosol jet printer providing in-flight aerosol characterization |
Citations (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6202734B1 (en) * | 1998-08-03 | 2001-03-20 | Sandia Corporation | Apparatus for jet application of molten metal droplets for manufacture of metal parts |
US20020171827A1 (en) * | 2001-05-17 | 2002-11-21 | Van Den Engh Ger | Apparatus for analyzing and sorting biological particles |
US6589792B1 (en) * | 1998-02-27 | 2003-07-08 | Dakocytomation Denmark A/S | Method and apparatus for flow cytometry |
US20040086159A1 (en) * | 2002-11-01 | 2004-05-06 | Lary Todd P. | Monitoring and control of droplet sorting |
US7159752B2 (en) * | 1997-12-12 | 2007-01-09 | Micron Technology, Inc. | Continuous mode solder jet apparatus |
US20080024619A1 (en) * | 2006-07-27 | 2008-01-31 | Hiroaki Ono | Image Processing Apparatus, Image Processing Method and Program |
US20080067068A1 (en) * | 2006-09-19 | 2008-03-20 | Vanderbilt University | DC-dielectrophoresis microfluidic apparatus, and applications of same |
US20100009445A1 (en) * | 2006-08-14 | 2010-01-14 | Mayo Foundation For Medical Education And Research | Rare earth nanoparticles |
US20100118300A1 (en) * | 2008-11-04 | 2010-05-13 | The Johns Hopkins University | Cylindrical illumination confocal spectroscopy system |
US20110275052A1 (en) * | 2002-08-01 | 2011-11-10 | Xy, Llc | Heterogeneous inseminate system |
US20110287976A1 (en) * | 2009-03-02 | 2011-11-24 | Jeff Tza-Huei Wang | Microfluidic solution for high-throughput, droplet-based single molecule analysis with low reagent consumption |
US20120076349A1 (en) * | 2009-06-03 | 2012-03-29 | Hitachi High-Technologies Corporation | Flow type particle image analysis method and device |
US20120135874A1 (en) * | 2009-05-08 | 2012-05-31 | The Johns Hopkins University | Single molecule spectroscopy for analysis of cell-free nucleic acid biomarkers |
US20120301869A1 (en) * | 2011-05-25 | 2012-11-29 | Inguran, Llc | Particle separation devices, methods and systems |
Family Cites Families (130)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3380584A (en) | 1965-06-04 | 1968-04-30 | Atomic Energy Commission Usa | Particle separator |
BE793185A (en) | 1971-12-23 | 1973-04-16 | Atomic Energy Commission | APPARATUS FOR QUICKLY ANALYZING AND SORTING PARTICLES SUCH AS BIOLOGICAL CELLS |
US3826364A (en) | 1972-05-22 | 1974-07-30 | Univ Leland Stanford Junior | Particle sorting method and apparatus |
US4009435A (en) * | 1973-10-19 | 1977-02-22 | Coulter Electronics, Inc. | Apparatus for preservation and identification of particles analyzed by flow-through apparatus |
US3924947A (en) | 1973-10-19 | 1975-12-09 | Coulter Electronics | Apparatus for preservation and identification of particles analyzed by flow-through apparatus |
DE2632962C3 (en) | 1976-07-22 | 1980-08-21 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V., 3400 Goettingen | Particle separator |
US4173415A (en) | 1976-08-20 | 1979-11-06 | Science Spectrum, Inc. | Apparatus and process for rapidly characterizing and differentiating large organic cells |
US4325483A (en) | 1979-08-20 | 1982-04-20 | Ortho Diagnostics, Inc. | Method for detecting and controlling flow rates of the droplet forming stream of an electrostatic particle sorting apparatus |
US4318480A (en) | 1979-08-20 | 1982-03-09 | Ortho Diagnostics, Inc. | Method and apparatus for positioning the point of droplet formation in the jetting fluid of an electrostatic sorting device |
US4318481A (en) | 1979-08-20 | 1982-03-09 | Ortho Diagnostics, Inc. | Method for automatically setting the correct phase of the charge pulses in an electrostatic flow sorter |
JPS5630870A (en) | 1979-08-23 | 1981-03-28 | Fuji Xerox Co Ltd | Ink jet printer |
US4284496A (en) | 1979-12-10 | 1981-08-18 | Newton William A | Particle guiding apparatus and method |
JPS58187441U (en) | 1982-06-09 | 1983-12-13 | 横河電機株式会社 | inkjet printer |
US4538733A (en) | 1983-10-14 | 1985-09-03 | Becton, Dickinson And Company | Particle sorter with neutralized collection wells and method of using same |
JPS6236542A (en) | 1985-08-09 | 1987-02-17 | Canon Inc | Particle analyzer |
US4616234A (en) | 1985-08-15 | 1986-10-07 | Eastman Kodak Company | Simultaneous phase detection and adjustment of multi-jet printer |
JPS62167478A (en) | 1985-11-29 | 1987-07-23 | Shimadzu Corp | Apparatus for dividedly taking particle |
JPS6412245A (en) | 1987-07-03 | 1989-01-17 | Canon Kk | Particle analyzing device |
US4987539A (en) | 1987-08-05 | 1991-01-22 | Stanford University | Apparatus and method for multidimensional characterization of objects in real time |
US5080770A (en) | 1989-09-11 | 1992-01-14 | Culkin Joseph B | Apparatus and method for separating particles |
EP0422616B1 (en) | 1989-10-11 | 1996-02-07 | Canon Kabushiki Kaisha | Apparatus for and method of fractionating particle in particle-suspended liquid in conformity with the properties thereof |
US5483469A (en) | 1993-08-02 | 1996-01-09 | The Regents Of The University Of California | Multiple sort flow cytometer |
US5700692A (en) * | 1994-09-27 | 1997-12-23 | Becton Dickinson And Company | Flow sorter with video-regulated droplet spacing |
US6861265B1 (en) | 1994-10-14 | 2005-03-01 | University Of Washington | Flow cytometer droplet formation system |
US5602039A (en) | 1994-10-14 | 1997-02-11 | The University Of Washington | Flow cytometer jet monitor system |
US5641457A (en) | 1995-04-25 | 1997-06-24 | Systemix | Sterile flow cytometer and sorter with mechanical isolation between flow chamber and sterile enclosure |
US5617911A (en) | 1995-09-08 | 1997-04-08 | Aeroquip Corporation | Method and apparatus for creating a free-form three-dimensional article using a layer-by-layer deposition of a support material and a deposition material |
DE19549015C1 (en) | 1995-12-28 | 1997-04-03 | Siemens Ag | Method of monitoring precise location at which fluid stream breaks up into droplets |
JP3258889B2 (en) | 1996-01-11 | 2002-02-18 | 株式会社堀場製作所 | Optical axis adjustment method in scattering particle size distribution analyzer |
JP2985826B2 (en) | 1997-04-09 | 1999-12-06 | 日本電気株式会社 | Position detecting apparatus and method |
US6079836A (en) | 1998-07-20 | 2000-06-27 | Coulter International Corp. | Flow cytometer droplet break-off location adjustment mechanism |
US6221254B1 (en) | 1998-08-25 | 2001-04-24 | J. Rodney Dickerson | Purification of liquid streams using carbon dioxide |
US6410872B2 (en) | 1999-03-26 | 2002-06-25 | Key Technology, Inc. | Agricultural article inspection apparatus and method employing spectral manipulation to enhance detection contrast ratio |
US6372506B1 (en) | 1999-07-02 | 2002-04-16 | Becton, Dickinson And Company | Apparatus and method for verifying drop delay in a flow cytometer |
US6813017B1 (en) * | 1999-10-20 | 2004-11-02 | Becton, Dickinson And Company | Apparatus and method employing incoherent light emitting semiconductor devices as particle detection light sources in a flow cytometer |
US7024316B1 (en) | 1999-10-21 | 2006-04-04 | Dakocytomation Colorado, Inc. | Transiently dynamic flow cytometer analysis system |
JP2001272328A (en) * | 2000-03-23 | 2001-10-05 | Sysmex Corp | Particle measurement apparatus by means of image |
US6583865B2 (en) * | 2000-08-25 | 2003-06-24 | Amnis Corporation | Alternative detector configuration and mode of operation of a time delay integration particle analyzer |
US7258774B2 (en) | 2000-10-03 | 2007-08-21 | California Institute Of Technology | Microfluidic devices and methods of use |
US7907765B2 (en) * | 2001-03-28 | 2011-03-15 | University Of Washington | Focal plane tracking for optical microtomography |
WO2002092247A1 (en) * | 2001-05-17 | 2002-11-21 | Cytomation, Inc. | Flow cytometer with active automated optical alignment system |
US7280207B2 (en) | 2001-07-25 | 2007-10-09 | Applera Corporation | Time-delay integration in a flow cytometry system |
US6949715B2 (en) | 2002-02-08 | 2005-09-27 | Kelly Arnold J | Method and apparatus for particle size separation |
US6866370B2 (en) | 2002-05-28 | 2005-03-15 | Eastman Kodak Company | Apparatus and method for improving gas flow uniformity in a continuous stream ink jet printer |
JP4099822B2 (en) | 2002-07-26 | 2008-06-11 | セイコーエプソン株式会社 | Dispensing device, dispensing method, and biological sample-containing solution ejection failure detection method |
US7201875B2 (en) | 2002-09-27 | 2007-04-10 | Becton Dickinson And Company | Fixed mounted sorting cuvette with user replaceable nozzle |
JP3979304B2 (en) | 2003-02-24 | 2007-09-19 | 日本光電工業株式会社 | Flow cell positioning method and flow cytometer with adjustable flow cell position |
JP4614947B2 (en) | 2003-03-28 | 2011-01-19 | イングラン・リミテッド・ライアビリティ・カンパニー | Apparatus and method for sorting particles and providing sex-sorted animal sperm |
ES2541121T3 (en) | 2003-05-15 | 2015-07-16 | Xy, Llc | Efficient classification of haploid cells by flow cytometry systems |
JP3875653B2 (en) * | 2003-06-05 | 2007-01-31 | 正昭 川橋 | Droplet state measuring device and state measuring method |
US7425253B2 (en) | 2004-01-29 | 2008-09-16 | Massachusetts Institute Of Technology | Microscale sorting cytometer |
US7232687B2 (en) | 2004-04-07 | 2007-06-19 | Beckman Coulter, Inc. | Multiple sorter monitor and control subsystem for flow cytometer |
EP1735428A4 (en) | 2004-04-12 | 2010-11-10 | Univ California | Optoelectronic tweezers for microparticle and cell manipulation |
JP4304120B2 (en) | 2004-04-30 | 2009-07-29 | ベイバイオサイエンス株式会社 | Apparatus and method for sorting biological particles |
PT1771729E (en) | 2004-07-27 | 2015-12-31 | Beckman Coulter Inc | Enhancing flow cytometry discrimination with geometric transformation |
US7410233B2 (en) | 2004-12-10 | 2008-08-12 | Konica Minolta Holdings, Inc. | Liquid droplet ejecting apparatus and a method of driving a liquid droplet ejecting head |
JP4047336B2 (en) | 2005-02-08 | 2008-02-13 | 独立行政法人科学技術振興機構 | Cell sorter chip with gel electrode |
JP4540506B2 (en) | 2005-03-04 | 2010-09-08 | 三井造船株式会社 | Method and apparatus for controlling position of sample liquid flow |
US7403125B2 (en) | 2005-05-06 | 2008-07-22 | Accuri Cytometers, Inc. | Flow cytometry system with bubble detection |
US7518108B2 (en) | 2005-11-10 | 2009-04-14 | Wisconsin Alumni Research Foundation | Electrospray ionization ion source with tunable charge reduction |
US7901947B2 (en) | 2006-04-18 | 2011-03-08 | Advanced Liquid Logic, Inc. | Droplet-based particle sorting |
JP4304195B2 (en) | 2006-06-13 | 2009-07-29 | ベイバイオサイエンス株式会社 | Apparatus and method for sorting biological particles |
US20070291058A1 (en) | 2006-06-20 | 2007-12-20 | Fagerquist Randy L | Continuous ink jet printing with satellite droplets |
JP4304634B2 (en) | 2006-10-23 | 2009-07-29 | ソニー株式会社 | Label detection apparatus and label detection method |
US7788969B2 (en) | 2006-11-28 | 2010-09-07 | Cummins Filtration Ip, Inc. | Combination contaminant size and nature sensing system and method for diagnosing contamination issues in fluids |
DE102006056694B4 (en) | 2006-11-30 | 2010-08-05 | Advalytix Ag | Method for carrying out an enzymatic reaction |
US8290625B2 (en) | 2007-04-04 | 2012-10-16 | Beckman Coulter, Inc. | Flow cytometer sorter |
WO2008130623A1 (en) | 2007-04-19 | 2008-10-30 | Brandeis University | Manipulation of fluids, fluid components and reactions in microfluidic systems |
US7828420B2 (en) | 2007-05-16 | 2010-11-09 | Eastman Kodak Company | Continuous ink jet printer with modified actuator activation waveform |
US7691636B2 (en) | 2007-05-23 | 2010-04-06 | Beckman Coulter, Inc. | Method and apparatus for compensating for variations in particle trajectories in electrostatic sorter for flowcell cytometer |
WO2009009081A2 (en) * | 2007-07-10 | 2009-01-15 | Massachusetts Institute Of Technology | Tomographic phase microscopy |
US7880108B2 (en) | 2007-10-26 | 2011-02-01 | Becton, Dickinson And Company | Deflection plate |
JP4990746B2 (en) | 2007-12-14 | 2012-08-01 | ベイバイオサイエンス株式会社 | Apparatus and method for separating biological particles contained in a liquid flow |
US9797010B2 (en) | 2007-12-21 | 2017-10-24 | President And Fellows Of Harvard College | Systems and methods for nucleic acid sequencing |
JP2009298012A (en) * | 2008-06-13 | 2009-12-24 | Konica Minolta Holdings Inc | Apparatus and method for inspecting discharge of liquid droplet, and image forming device |
JP4572973B2 (en) | 2008-06-16 | 2010-11-04 | ソニー株式会社 | Microchip and flow-feeding method in microchip |
NZ590166A (en) * | 2008-06-30 | 2013-09-27 | Microbix Biosystems Inc | Method and apparatus for sorting cells |
JP5487638B2 (en) | 2009-02-17 | 2014-05-07 | ソニー株式会社 | Apparatus for microparticle sorting and microchip |
JP5078929B2 (en) | 2009-03-17 | 2012-11-21 | 三井造船株式会社 | Cell sorter and sample separation method |
EP3415235A1 (en) | 2009-03-23 | 2018-12-19 | Raindance Technologies Inc. | Manipulation of microfluidic droplets |
JP5304456B2 (en) | 2009-06-10 | 2013-10-02 | ソニー株式会社 | Fine particle measuring device |
JP5321260B2 (en) | 2009-06-11 | 2013-10-23 | ソニー株式会社 | Optical measuring device, flow cytometer and optical measuring method |
US8628648B2 (en) | 2009-07-07 | 2014-01-14 | The University Of Akron | Apparatus and method for manipulating micro component |
JP5446563B2 (en) * | 2009-08-06 | 2014-03-19 | ソニー株式会社 | Fine particle sorting device and flow cytometer using the fine particle sorting device |
US8570511B2 (en) | 2009-09-09 | 2013-10-29 | Brookhaven Science Associates, Llc | Wide size range fast integrated mobility spectrometer |
US9151646B2 (en) | 2011-12-21 | 2015-10-06 | Deka Products Limited Partnership | System, method, and apparatus for monitoring, regulating, or controlling fluid flow |
EP2545356B1 (en) | 2010-03-09 | 2015-12-02 | Beckman Coulter, Inc. | Calculate drop delay for flow cytometry systems and methods |
CN102272580B (en) | 2010-03-31 | 2014-07-30 | 古河电气工业株式会社 | Optical information analysis device and optical information analysis method |
JP5437148B2 (en) | 2010-04-23 | 2014-03-12 | ベイバイオサイエンス株式会社 | Flow cytometer and cell sorter |
JP2011237201A (en) | 2010-05-06 | 2011-11-24 | Sony Corp | Particulate dispensing device, microchip, and microchip module |
US8922636B1 (en) * | 2010-08-20 | 2014-12-30 | The United States Of America As Represented By The Secretary Of The Navy | Synthetic aperture imaging for fluid flows |
JP2012047464A (en) | 2010-08-24 | 2012-03-08 | Sony Corp | Fine particle measuring instrument and optical axis correction method |
US9170138B2 (en) | 2010-10-01 | 2015-10-27 | The Board Of Trustees Of The Leland Stanford Junior University | Enhanced microfluidic electromagnetic measurements |
CA2826544C (en) | 2011-02-04 | 2020-06-30 | Cytonome/St, Llc | Particle sorting apparatus and method |
US9267873B2 (en) | 2011-03-30 | 2016-02-23 | Empire Technology Development Llc | Material sorting system and method of sorting material |
KR101569600B1 (en) | 2011-06-08 | 2015-11-16 | 엠파이어 테크놀로지 디벨롭먼트 엘엘씨 | Two-dimensional image capture for an augmented reality representation |
JP6003020B2 (en) | 2011-08-03 | 2016-10-05 | ソニー株式会社 | Microchip and fine particle analyzer |
US9897530B2 (en) | 2011-08-25 | 2018-02-20 | Sony Corporation | Compensation of motion-related error in a stream of moving micro-entities |
JP5880088B2 (en) | 2012-01-31 | 2016-03-08 | ブラザー工業株式会社 | Edge detection apparatus, image data processing apparatus, liquid ejection apparatus including the image data processing apparatus, edge detection method, and edge detection program |
US9324190B2 (en) | 2012-02-24 | 2016-04-26 | Matterport, Inc. | Capturing and aligning three-dimensional scenes |
US20150132766A1 (en) * | 2012-03-30 | 2015-05-14 | On-Chip Cellomics Consortium | Imaging cell sorter |
JP5924077B2 (en) | 2012-03-30 | 2016-05-25 | ソニー株式会社 | Fine particle sorting device and method for determining orbit direction in fine particle sorting device |
JPWO2013145836A1 (en) | 2012-03-30 | 2015-12-10 | ソニー株式会社 | Microchip type optical measuring apparatus and optical position adjusting method in the apparatus |
JP5994337B2 (en) | 2012-03-30 | 2016-09-21 | ソニー株式会社 | Fine particle sorting device and delay time determination method |
WO2013145905A1 (en) | 2012-03-30 | 2013-10-03 | ソニー株式会社 | Microparticle fractionation apparatus, and method for optimizing fluid stream in said apparatus |
EP3511694A1 (en) | 2012-03-30 | 2019-07-17 | Sony Corporation | Microparticle sorting device and method for controlling position in microparticle sorting device |
JP5924276B2 (en) | 2012-04-03 | 2016-05-25 | ソニー株式会社 | Channel device, particle sorting apparatus, and particle sorting method |
US20130286038A1 (en) | 2012-04-30 | 2013-10-31 | General Electric Company | Systems and methods for selection and display of multiplexed images of biological tissue |
JP2014020918A (en) | 2012-07-18 | 2014-02-03 | Sony Corp | Microparticle measuring instrument and microparticle analysis method |
WO2014017186A1 (en) | 2012-07-25 | 2014-01-30 | ソニー株式会社 | Microparticle measurement device and liquid supply method for microparticle measurement device |
US9168568B2 (en) | 2012-08-01 | 2015-10-27 | Owl biomedical, Inc. | Particle manipulation system with cytometric confirmation |
JP2014062822A (en) | 2012-09-21 | 2014-04-10 | Sony Corp | Fine particle analyzer and fine particle analyzing method |
JP6065527B2 (en) | 2012-11-08 | 2017-01-25 | ソニー株式会社 | Fine particle sorting device and fine particle sorting method |
WO2014115409A1 (en) | 2013-01-28 | 2014-07-31 | ソニー株式会社 | Fine particle fractionation device, fine particle fractionation method and program |
US9915599B2 (en) | 2013-02-08 | 2018-03-13 | Sony Corporation | Microparticle analysis apparatus and microparticle analysis system |
JP2014174139A (en) | 2013-03-13 | 2014-09-22 | Sony Corp | Flow channel device, particle sorter, particle outflow method, and particle sorting method |
EP2972206B1 (en) * | 2013-03-14 | 2024-02-21 | Cytonome/ST, LLC | Operatorless particle processing systems and methods |
EP2984468B1 (en) | 2013-04-12 | 2021-11-17 | Becton, Dickinson and Company | Automated set-up for cell sorting |
US9645080B2 (en) | 2013-04-16 | 2017-05-09 | University Of Washington | Systems, devices, and methods for separating, concentrating, and/or differentiating between cells from a cell sample |
EP3004813A4 (en) | 2013-05-29 | 2016-12-21 | Gnubio Inc | Low cost optical high speed discrete measurement system |
US10309891B2 (en) | 2013-10-16 | 2019-06-04 | Sony Corporation | Particle sorting apparatus, particle sorting method, and program |
JP6136843B2 (en) | 2013-10-17 | 2017-05-31 | ソニー株式会社 | Particle sorting apparatus, particle sorting method and program |
JP6465036B2 (en) | 2014-02-13 | 2019-02-06 | ソニー株式会社 | Particle sorting device, particle sorting method, program, and particle sorting system |
JP6102783B2 (en) | 2014-02-14 | 2017-03-29 | ソニー株式会社 | Particle sorting apparatus, particle sorting method and program |
JP6657625B2 (en) | 2014-09-05 | 2020-03-04 | ソニー株式会社 | Droplet sorting apparatus, drop sorting method and program |
CN106663411A (en) | 2014-11-16 | 2017-05-10 | 易欧耐特感知公司 | Systems and methods for augmented reality preparation, processing, and application |
CN108139312B (en) | 2015-10-19 | 2021-02-05 | 索尼公司 | Image processing apparatus, microparticle sorting apparatus, and image processing method |
WO2017073737A1 (en) | 2015-10-28 | 2017-05-04 | 国立大学法人東京大学 | Analysis device |
WO2018067210A1 (en) | 2016-10-03 | 2018-04-12 | Becton, Dickinson And Company | Methods and systems for determining a drop delay of a flow stream in a flow cytometer |
US10466158B2 (en) | 2017-04-11 | 2019-11-05 | Sony Corporation | Microparticle sorting apparatus and delay time determination method |
-
2012
- 2012-03-30 JP JP2012080366A patent/JP5924077B2/en active Active
-
2013
- 2013-03-07 US US13/788,165 patent/US20130258075A1/en not_active Abandoned
- 2013-03-22 CN CN201310095425.0A patent/CN103364325B/en active Active
-
2019
- 2019-07-19 US US16/517,281 patent/US11193874B2/en active Active
Patent Citations (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7159752B2 (en) * | 1997-12-12 | 2007-01-09 | Micron Technology, Inc. | Continuous mode solder jet apparatus |
US6589792B1 (en) * | 1998-02-27 | 2003-07-08 | Dakocytomation Denmark A/S | Method and apparatus for flow cytometry |
US6202734B1 (en) * | 1998-08-03 | 2001-03-20 | Sandia Corporation | Apparatus for jet application of molten metal droplets for manufacture of metal parts |
US20020171827A1 (en) * | 2001-05-17 | 2002-11-21 | Van Den Engh Ger | Apparatus for analyzing and sorting biological particles |
US20110275052A1 (en) * | 2002-08-01 | 2011-11-10 | Xy, Llc | Heterogeneous inseminate system |
US20040086159A1 (en) * | 2002-11-01 | 2004-05-06 | Lary Todd P. | Monitoring and control of droplet sorting |
US20080024619A1 (en) * | 2006-07-27 | 2008-01-31 | Hiroaki Ono | Image Processing Apparatus, Image Processing Method and Program |
US20100009445A1 (en) * | 2006-08-14 | 2010-01-14 | Mayo Foundation For Medical Education And Research | Rare earth nanoparticles |
US20080067068A1 (en) * | 2006-09-19 | 2008-03-20 | Vanderbilt University | DC-dielectrophoresis microfluidic apparatus, and applications of same |
US20100118300A1 (en) * | 2008-11-04 | 2010-05-13 | The Johns Hopkins University | Cylindrical illumination confocal spectroscopy system |
US20110287976A1 (en) * | 2009-03-02 | 2011-11-24 | Jeff Tza-Huei Wang | Microfluidic solution for high-throughput, droplet-based single molecule analysis with low reagent consumption |
US20120135874A1 (en) * | 2009-05-08 | 2012-05-31 | The Johns Hopkins University | Single molecule spectroscopy for analysis of cell-free nucleic acid biomarkers |
US20120076349A1 (en) * | 2009-06-03 | 2012-03-29 | Hitachi High-Technologies Corporation | Flow type particle image analysis method and device |
US20120301869A1 (en) * | 2011-05-25 | 2012-11-29 | Inguran, Llc | Particle separation devices, methods and systems |
Cited By (63)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8831306B2 (en) * | 2009-06-03 | 2014-09-09 | Hitachi High-Technologies Corporation | Flow type particle image analysis method and device |
US20120076349A1 (en) * | 2009-06-03 | 2012-03-29 | Hitachi High-Technologies Corporation | Flow type particle image analysis method and device |
US9255874B2 (en) * | 2011-02-04 | 2016-02-09 | Cytonome/St, Llc | Fluid stream imaging apparatus |
US20140176704A1 (en) * | 2011-02-04 | 2014-06-26 | Cytonome/St, Llc | Fluid stream imaging apparatus |
US9958375B2 (en) | 2012-03-30 | 2018-05-01 | Sony Corporation | Microparticle sorting apparatus and delay time determination method |
US9087371B2 (en) | 2012-03-30 | 2015-07-21 | Sony Corporation | Microparticle sorting device and method of optimizing fluid stream therein |
US10132735B2 (en) | 2012-03-30 | 2018-11-20 | Sony Corporation | Microparticle sorting device and method of optimizing fluid stream therein |
US9339823B2 (en) | 2012-03-30 | 2016-05-17 | Sony Corporation | Microparticle sorting apparatus and delay time determination method |
US10859996B2 (en) | 2012-03-30 | 2020-12-08 | Sony Corporation | Microchip-type optical measuring apparatus and optical position adjusting method thereof |
US9029724B2 (en) | 2012-03-30 | 2015-05-12 | Sony Corporation | Microparticle sorting device and method for controlling position in microparticle sorting device |
US11193874B2 (en) | 2012-03-30 | 2021-12-07 | Sony Corporation | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles |
US9915935B2 (en) | 2012-03-30 | 2018-03-13 | Sony Corporation | Microchip-type optical measuring apparatus and optical position adjusting method thereof |
US10876954B2 (en) | 2012-03-30 | 2020-12-29 | Sony Corporation | Microparticle sorting apparatus and delay time determination method |
US10180676B2 (en) | 2012-03-30 | 2019-01-15 | Sony Corporation | Microchip-type optical measuring apparatus and optical position adjusting method thereof |
US10646866B2 (en) | 2012-07-25 | 2020-05-12 | Sony Corporation | Microparticle measurement device and liquid delivery method in microparticle measurement device |
US9669403B2 (en) | 2012-07-25 | 2017-06-06 | Sony Corporation | Microparticle measurement device and liquid delivery method in microparticle measurement device |
US9784659B2 (en) | 2012-11-08 | 2017-10-10 | Sony Corporation | Microparticle fractionating apparatus and method of fractionating microparticle |
US11313784B2 (en) | 2013-01-28 | 2022-04-26 | Sony Corporation | Microparticle sorting device, and method and program for sorting microparticles |
US9784660B2 (en) | 2013-01-28 | 2017-10-10 | Sony Corporation | Microparticle sorting device, and method and program for sorting microparticles |
US10241025B2 (en) | 2013-01-28 | 2019-03-26 | Sony Corporation | Microparticle sorting device, and method and program for sorting microparticles |
US9915599B2 (en) | 2013-02-08 | 2018-03-13 | Sony Corporation | Microparticle analysis apparatus and microparticle analysis system |
US9964968B2 (en) | 2013-03-14 | 2018-05-08 | Cytonome/St, Llc | Operatorless particle processing systems and methods |
US10871790B2 (en) | 2013-03-14 | 2020-12-22 | Cytonome/St, Llc | Operatorless particle processing systems and methods |
US8975595B2 (en) * | 2013-04-12 | 2015-03-10 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US11060894B2 (en) | 2013-04-12 | 2021-07-13 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US9952076B2 (en) | 2013-04-12 | 2018-04-24 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US9366616B2 (en) | 2013-04-12 | 2016-06-14 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US9726527B2 (en) | 2013-04-12 | 2017-08-08 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US10578469B2 (en) | 2013-04-12 | 2020-03-03 | Becton, Dickinson And Company | Automated set-up for cell sorting |
US20140306122A1 (en) * | 2013-04-12 | 2014-10-16 | Becton, Dickinson And Company | Automated set-up for cell sorting |
EP3035030A4 (en) * | 2013-10-16 | 2017-03-22 | Sony Corporation | Particle fractionation device, particle fractionation method, and program |
CN105659069A (en) * | 2013-10-16 | 2016-06-08 | 索尼公司 | Particle fractionation device, particle fractionation method, and program |
US10309891B2 (en) | 2013-10-16 | 2019-06-04 | Sony Corporation | Particle sorting apparatus, particle sorting method, and program |
US9857286B2 (en) | 2013-10-17 | 2018-01-02 | Sony Corporation | Particle fractionation apparatus, particle fractionation method and particle fractionation program |
EP3690424A1 (en) * | 2014-02-13 | 2020-08-05 | Sony Corporation | Particle sorting device, particle sorting method, program, and particle sorting system |
CN105980831A (en) * | 2014-02-13 | 2016-09-28 | 索尼公司 | Particle sorting apparatus, particle sorting method, program, and particle sorting system |
US20190323945A1 (en) * | 2014-02-13 | 2019-10-24 | Sony Corporation | Particle sorting device, particle sorting method, program, and particle sorting system |
US11119030B2 (en) | 2014-02-13 | 2021-09-14 | Sony Corporation | Particle sorting device, particle sorting method, program, and particle sorting system |
US10309892B2 (en) | 2014-02-13 | 2019-06-04 | Sony Corporation | Particle sorting device, particle sorting method, program, and particle sorting system |
EP3106857A4 (en) * | 2014-02-13 | 2017-10-18 | Sony Corporation | Particle sorting apparatus, particle sorting method, program, and particle sorting system |
US20180045638A1 (en) * | 2014-02-14 | 2018-02-15 | Sony Corporation | Particle sorting apparatus, particle sorting method, and non-transitory computer-readable storage medium storing program |
US20170010203A1 (en) * | 2014-02-14 | 2017-01-12 | Sony Corporation | Particle sorting apparatus, particle sorting method, and non-transitory computer-readable storage medium storing program |
US10451534B2 (en) * | 2014-02-14 | 2019-10-22 | Sony Corporation | Particle sorting apparatus and particle sorting method |
US9804075B2 (en) * | 2014-02-14 | 2017-10-31 | Sony Corporation | Particle sorting apparatus, particle sorting method, and non-transitory computer-readable storage medium storing program |
US20170074775A1 (en) * | 2014-05-22 | 2017-03-16 | Sony Corporation | Particle analyzer |
US10006849B2 (en) * | 2014-05-22 | 2018-06-26 | Sony Corporation | Particle analyzer |
US10156510B2 (en) | 2014-08-28 | 2018-12-18 | Sysmex Corporation | Particle imaging apparatus and particle imaging method |
US10386287B2 (en) | 2014-09-05 | 2019-08-20 | Sony Corporation | Droplet sorting device, droplet sorting method and program |
US10876952B2 (en) | 2014-09-05 | 2020-12-29 | Sony Corporation | Droplet sorting device, droplet sorting method and program |
US10723497B2 (en) * | 2014-11-03 | 2020-07-28 | Vanrx Pharmasystems Inc. | Apparatus and method for monitoring and controlling the filling of a container with a pharmaceutical fluid in an aseptic environment |
US10605714B2 (en) | 2015-10-19 | 2020-03-31 | Sony Corporation | Image processing device, fine particle sorting device, and image processing method |
US11204309B2 (en) | 2015-10-19 | 2021-12-21 | Sony Corporation | Image processing device, fine particle sorting device, and image processing method |
WO2017222461A1 (en) * | 2016-06-21 | 2017-12-28 | Giatrellis Sarantis | System and method for precision deposition of liquid droplets |
EP3511691A4 (en) * | 2016-09-12 | 2019-12-18 | Sony Corporation | Microparticle measurement device and microparticle measurement method |
WO2019008044A1 (en) * | 2017-07-04 | 2019-01-10 | Cytena Gmbh | Method for dispensing a liquid |
CN112400103A (en) * | 2018-07-09 | 2021-02-23 | 生德奈股份有限公司 | Method and dispensing device for examining a liquid sample |
WO2020011773A1 (en) * | 2018-07-09 | 2020-01-16 | Cytena Gmbh | Method for examining a liquid sample and a dispensing apparatus |
AU2019301870B2 (en) * | 2018-07-09 | 2022-06-23 | Cytena Gmbh | Method for examining a liquid sample and a dispensing apparatus |
US20210270721A1 (en) * | 2018-10-01 | 2021-09-02 | Hewlett-Packard Development Company, L.P. | Particle sorting using microfluidic ejectors |
US11486814B2 (en) * | 2018-10-01 | 2022-11-01 | Hewlett-Packard Development Company, L.P. | Particle sorting using microfluidic ejectors |
WO2021262359A1 (en) * | 2020-06-24 | 2021-12-30 | Becton, Dickinson And Company | Flow cytometric droplet dispensing systems and methods for using the same |
US11754487B2 (en) | 2020-06-24 | 2023-09-12 | Becton, Dickinson And Company | Flow cytometric droplet dispensing systems and methods for using the same |
CN115414972A (en) * | 2022-08-08 | 2022-12-02 | 广东省科学院生物与医学工程研究所 | Portable coaxial focusing micro-droplet generation device and micro-droplet preparation method |
Also Published As
Publication number | Publication date |
---|---|
JP5924077B2 (en) | 2016-05-25 |
US11193874B2 (en) | 2021-12-07 |
US20200011783A1 (en) | 2020-01-09 |
JP2013210270A (en) | 2013-10-10 |
CN103364325B (en) | 2017-09-08 |
CN103364325A (en) | 2013-10-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11193874B2 (en) | Micro-particle sorting apparatus and method of determining a trajectory of an ejected stream carrying micro-particles | |
US10876954B2 (en) | Microparticle sorting apparatus and delay time determination method | |
US11204309B2 (en) | Image processing device, fine particle sorting device, and image processing method | |
JP6102994B2 (en) | Fine particle sorting device and position control method in fine particle sorting device | |
JP6256537B2 (en) | Microchip type optical measuring apparatus and optical position adjusting method in the apparatus | |
KR102318759B1 (en) | Fine particle sorting device and method of determining delay time | |
JP6465036B2 (en) | Particle sorting device, particle sorting method, program, and particle sorting system | |
US8941081B2 (en) | Microparticle measurement apparatus and microparticle analysis method | |
CN105980059A (en) | Particle sorting apparatus, particle sorting method, and non-transitory computer-readable storage medium storing program | |
WO2014017186A1 (en) | Microparticle measurement device and liquid supply method for microparticle measurement device | |
JP2013210287A (en) | Calibration method in microparticle separation device, the device and calibration particle | |
JP6237806B2 (en) | Fine particle fractionator | |
JP6135796B2 (en) | Fine particle sorting device and method for determining orbit direction in fine particle sorting device |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: SONY CORPORATION, JAPAN Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MURAKI, YOSUKE;TSUJI, AKIKO;SIGNING DATES FROM 20130131 TO 20130304;REEL/FRAME:029968/0596 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |