US20010028458A1 - Compact spectrofluorometer - Google Patents

Compact spectrofluorometer Download PDF

Info

Publication number
US20010028458A1
US20010028458A1 US09/813,325 US81332501A US2001028458A1 US 20010028458 A1 US20010028458 A1 US 20010028458A1 US 81332501 A US81332501 A US 81332501A US 2001028458 A1 US2001028458 A1 US 2001028458A1
Authority
US
United States
Prior art keywords
filter
light
active
area
instrument
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.)
Granted
Application number
US09/813,325
Other versions
US6441892B2 (en
Inventor
Ming Xiao
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Horiba Instruments Inc
Original Assignee
Horiba Jobin Yvon Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority claimed from US09/443,392 external-priority patent/US6323944B1/en
Application filed by Horiba Jobin Yvon Inc filed Critical Horiba Jobin Yvon Inc
Priority to US09/813,325 priority Critical patent/US6441892B2/en
Assigned to JOBIN YVON, INC. reassignment JOBIN YVON, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: XIAO, MING
Publication of US20010028458A1 publication Critical patent/US20010028458A1/en
Application granted granted Critical
Publication of US6441892B2 publication Critical patent/US6441892B2/en
Assigned to HORIBA JOBIN YVON INC. reassignment HORIBA JOBIN YVON INC. CHANGE OF NAME (SEE DOCUMENT FOR DETAILS). Assignors: JOBIN YVON INC.
Assigned to HORIBA INSTRUMENTS INCORPORATED reassignment HORIBA INSTRUMENTS INCORPORATED CHANGE OF NAME (SEE DOCUMENT FOR DETAILS). Assignors: HORIBA JOBIN YVON INC.
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01JMEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/12Generating the spectrum; Monochromators
    • G01J3/26Generating the spectrum; Monochromators using multiple reflection, e.g. Fabry-Perot interferometer, variable interference filters
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01JMEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02Details
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01JMEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02Details
    • G01J3/0294Multi-channel spectroscopy
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01JMEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/28Investigating the spectrum
    • G01J3/44Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
    • G01J3/4406Fluorescence spectrometry
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • G01N21/6456Spatial resolved fluorescence measurements; Imaging
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N2021/6417Spectrofluorimetric devices
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N2021/6417Spectrofluorimetric devices
    • G01N2021/6423Spectral mapping, video display

Definitions

  • Fluorescence instrumentation has been used for many years to identify unknown materials.
  • the principle involved is that a material excited with light of a particular wavelength will emit light energy in the form of an emission spectrum whose amplitude profile, over the range of wavelengths emitted, constitutes a “fingerprint” which can give the identity and nature of the unknown material.
  • FIG. 1 Such a prior art system is illustrated in FIG. 1. Measurement of the fluorescence spectrum is achieved by having a system which comprises an excitation spectrograph 1 which is used to excite a sample 2 , typically contained in an elongated cuvette 3 .
  • the elongated cuvette 3 is excited by an elongated image of a spectrum extending from a low wavelength to a high wavelength.
  • a xenon source is imaged as a bright line placed over a cuvette in a vertical line.
  • the full spectrum will excite any homogeneous sample placed in the sample compartment of the cuvette.
  • the resulting fluorescence emission is dispersed orthogonally over the active area of a rectangular CCD, or charge-coupled device, which is, essentially, a two-dimensional array of light detectors.
  • the horizontal axis of the CCD records the emission spectra at different excitation wavelengths along the vertical axis, and gives the intensity for each wavelength.
  • this instrument will produce, for each wavelength in the range of excitation wavelengths, the spectrum of emitted wavelengths. For example, if the system has a resolution of 5 nm, and covers a range of 100 nm, one could view the output as twenty different spectra.
  • a cuvette may be fed by a high pressure liquid chromatography column, allowing the facile real-time generation of fluorescence emission spectra of the various materials in a sample being analyzed by the chromatography column.
  • an excitation light source provides optical radiation over a range of wavelengths or spectra for illuminating a sample.
  • the inventive instrument performs fluoresence analysis of samples, and comprises a light source emitting light into an illumination light path, and a first spectral filter in the illumination light path for transmitting light within a selected wavelength range. This defines a sample illumination light path.
  • a second spectral filter is spaced from the first spectral filter forming a sample receiving space therebetween.
  • the illumination light path passes through the first spectral filter.
  • the sample receiver and the second spectral filter lie in the light path, and the second spectral filter is displaced angularly relative to the first spectral filter.
  • a sensing element in the resultant light path measures absorption spectra and fluorescence light.
  • the first spectral filter and the second spectral filter have a characteristic which varies along an axis thereof.
  • the variable characteristic is a variable bandpass wavelength in various filter regions of the spectral filter.
  • the second spectral filter is angularly displaced at a substantially othogonal angle.
  • the above described embodiment of the invention has the advantage of providing along a diagonal region of the CCD the absorption spectrum of the material sample under analysis.
  • a third spectral filter in the resultant light path is oriented in a direction, and position in a position which are substantially the same as the direction and position of the first spectral filter.
  • This third filter serves the function of a blocking filter thereby preventing excitation light energy that has passed through a sample receiver from passing to the sensing element or CCD array.
  • FIG. 1 is a schematic view of a typical prior art apparatus
  • FIG. 2 is a schematic view of the present invention showing the compactness of the components
  • FIG. 3 is an exploded schematic view of the apparatus of the present invention showing a pair of linear variable spectral filter and a CCD type of sensing element;
  • FIG. 4 is an exploded view similar to FIG. 3 including a cartridge containing a sample to be tested;
  • FIG. 5 is a schematic view of a CCD sensing element, as is employed in the embodiments illustrated in FIGS. 4 and 5;
  • FIG. 6 is a schematic view of a CCD sensing element, as is employed in the embodiments illustrated in FIGS. 4 and 5, illustrating the absorption spectrum position when filter elements are not matched in the system of the present invention
  • FIG. 7 is a perspective view of an alternative embodiment of the present invention.
  • FIG. 8 is a perspective view of an alternative embodiment of the inventive spectrofluorometer incorporating a further improvement
  • FIG. 9 is a view similar to FIG. 4 of an alternative embodiment of the invention including an excitation light blocking filter.
  • FIG. 10 is a perspective view of an embodiment of the invention similar to that illustrated in FIG. 9 and incorporating minimized light paths.
  • Optical radiation traveling along an excitation light path 12 passes into a linear variable spectral filter 14 .
  • Spectral filter 14 is a device which has bandpass wavelength characteristics which vary along its length. More particularly, at the bottom of filter 14 , one wavelength would be passed in the region defined by the dashed lines. In the next filter region above that filter region like having a different wavelength will be passed, perhaps a wavelength which is 5 nm longer.
  • This sort of device is made by advancing a mask having the width of one of the regions illustrated in dashed lines in the figure, from one discrete position to another and applying a different multilayer structure at each position to give the corresponding stripe of bandpass material the desired optical bandpass characteristic.
  • Such a filter has a spectral range of 400 to 700 nm. It is relatively small and compact, being 60 mm long, 25 mm wide and 5 mm thick. A typical spectrum length would be 44 mm, with dispersion varying between 0.12 and 0.17 mm/nm.
  • linear variable spectral filters sold by this corporation tend to vary in their characteristics, with a spectrum length varying form 37 to 51 mm. matching of the filters used in the embodiment of FIG. 2 is desirable.
  • a computer reading the output of the system may calibrate the software against a known source.
  • sample receiver 16 is located between the first spectral filter 14 and a second linear variable spectral filter 18 .
  • Sample receiver 16 is a vessel which defines a volume for receiving a sample which is to be analyzed. It may be a rectangular solid made of glass, plastic or any suitable material. It may also be as simple as a glass slide with a smear of the sample, or even a solid film of the sample material, such as tissue, paper from a paper mill whose operation is being monitored, and so forth.
  • Such a sample may be a solution derived from a material being tested, blood, the output of an HPLC liquid chromatography column, or the like. If the output of an HPLC column is being monitored, the receiver 16 may have a liquid input port and a drain, and the dimensions of the receiver would be such that capillary action insures the presence of sample material throughout the excited regions of receiver 16 .
  • a close-coupled discharge (CCD) sensing element 20 measures the relative position and intensity of light rays traveling along a resultant light path 12 . See FIG. 3.
  • Sensing element 20 is preferably a CCD type of sensor although other types can be used depending upon the type of excitation light used and the sample to be tested.
  • detector 20 is shown as a 36 element matrix detector.
  • the small number of elements or pixels is merely for the convenience of illustration and the illustration of the principles of the invention. In a real device, the number of detectors easily ranges into the hundreds of thousands of elements, and, depending upon the performances desired and the nature of the software reading out the signal from the detector, the number of elements in detector 20 may range into the millions of pixels.
  • First filter 14 is a linear variable spectral filter that changes its bandpass wavelength along the length or planar axis 15 of the filter. Wavelengths outside the desired transmission ranges are blocked by the respective filter regions.
  • the spectral range from 400 to 700 nm is oriented vertically, e.g., with shortest wavelength filter region 24 at the bottom, then longer wavelength filter region 26 , still longer wavelength filter region 28 , a filter region 30 which passes a range of wavelengths longer than those of filter region 28 , a filter region 32 which passes a range of wavelengths longer than those of filter region 30 , and the longest wavelength bandpass filter region than 34 at the top. While the invention has been implemented with a spectral filter having the aforementioned wavelength characteristics, other visible and non-visible bandpass characteristics can be used depending on the nature and characteristics of the sample to be tested.
  • the second optical filter 18 is substantially the same as the first optical filter 14 except that it is oriented in such a manner that its gradations are not in line with those of first filter 14 .
  • the strips defining the bandpass filter regions on filter 18 are preferably at ninety degrees to those of filter 14 .
  • a light source 36 which may comprise a xenon lamp whose output is collimated by a lens or reflector, or any other suitable optical components produces an excitation white light ray bundle 38 , sometimes referred to as illumination light, that travels along excitation light path 12 with a wide range of wavelengths striking the surface of filter 14 .
  • illumination light an excitation white light ray bundle 38
  • filter 14 As white light ray bundle 38 passes through filter 14 , selected wavelengths are passed by each filter region, such that a wavelength “gradient” from short to long wavelengths is produced. This is referred to herein as a sample excitation light 42 .
  • sample excitation light 42 passes through second filter 18 , only those wavelengths of light that are not blocked pass completely through the filter 18 . Since filter 18 is oriented at a right angle to filter 14 , most of sample excitation light 42 is blocked. By way of example, ⁇ 1 passes through filter 14 and filter 18 , while ⁇ 2 passes through filter 14 , but is blocked by filter 18 . In this manner a diagonal spectral line 56 is transmitted onto sensing element 20 . The theoretical center of this line it illustrated in FIG. 5 by phantom line 56 . This intrinsic relationship between the two linear variable spectral filters provides for simplicity of design, ruggedness and compact size of the inventive spectrofluorometer 10 .
  • Sample receiver 16 is located between filter 14 and filter 18 .
  • Sample receiver 16 may be any of a number of conventional sample holding types or techniques. As sample excitation light passes through sample 44 some of the light energy is converted into fluorescence emissions. The physics of this conversion are well understood and generally involve the photon of excitation radiation raising the energy level of electrons in the excited atom to a higher energy level or shell. When the electron snaps back into its unexcited state, it emits a photon with an energy level lower that the exciting photon, thus resulting in the fluorescence having a wavelength longer than the excitation wavelength.
  • sample excitation light is “absorbed” by sample 44 and does not contribute to the emission. The net result is to increase the kinetic energy of the atoms of the sample, and thus raise the temperature of the sample.
  • a resultant light ray bundle 50 exiting sample receiver 16 , comprises light rays which have exited filter 14 and fluoresence emissions from molecules that have been excited by light rays which have exited filter 14 .
  • Resultant light ray bundle 50 then passes into filter 18 where a selected wavelengths of both spectral light and fluorescent light are selectively blocked along the spectral gradient.
  • the portions of light ray bundle 50 passing through to sensing element 20 constitutes the absorption spectrum 52 of the material being analyzed and appears along imaginary line 56 in FIG. 5. This can be used to identify sample 44 .
  • filters 14 and 18 are substantially identical, but are positioned with their bandpass filter strip filter regions 24 - 34 and 35 - 44 oriented at right angles to each other.
  • filter region 24 has the same bandpass characteristic as filter region 34 .
  • filter region 26 has the same bandpass characteristic as filter region 42 .
  • Filter region 28 has the same bandpass characteristic as filter region 40 .
  • Filter region 30 has the same bandpass characteristic as filter region 37 .
  • Filter region 32 has the same bandpass characteristic as filter region 36 .
  • Filter region 34 has the same bandpass characteristic as filter region 35 .
  • the CCD elements 70 lying along line 56 in FIG. 5, are the only elements that will be illuminated by the white light ray bundle 38 coming from the excitation source. Moreover, because the fluorescence spectrum constitutes only wavelengths longer than the excitation wavelength, they will be blocked from reaching elements 70 by filter 18 . Thus, only the absorption spectrum can be seen along imaginary line 56 to provide a first identification of the sample.
  • the fluorescence spectrum constitutes only wavelengths longer than the excitation wavelength, these longer wavelengths will be passed by filter 18 to those elements 58 of the CCD which lie below line 56 in FIG. 5.
  • the elements 58 of the CCD which lie below line 56 in FIG. 5 produce the fluorescence emission spectra of the sample under analysis. The resultant fluorescence emission is used to identify sample 44 .
  • the operation of the inventive system may be better understood.
  • the output of the xenon lamp 36 constituting a broadband emission which is collimated into white light ray bundle 38 is caused to fall on filter 14 , which outputs a plurality of stripes of light energy at different wavelengths.
  • filters 14 and 18 are very thin, as is sample container 16 , the output of filter 14 is effectively “imaged” on the sample in sample receiver 16 .
  • the output of sample container 16 is likewise effectively “imaged” on filter 18 .
  • the output of filter 18 is effectively “imaged” on the surface of CCD elements 58 .
  • the system works because all of the above thin elements are in contact with each other and CCD 20 to form the sandwich illustrated in FIG. 2.
  • light ray 72 which is one of the light rays in white light bundle 38 , because it is in the bandpass range of filter region 34 on filter 14 , and, naturally, in the bandpass of optically identical filter region 35 , will pass through both filters and fall on CCD 20 , if it is not absorbed by the sample.
  • light ray 74 which is in the bandpass of filter regions 24 and 44 .
  • Light rays 76 and 78 will, on the other hand, be blocked by filter 18 , after being limited to the different bandpass of facing filter regions of filter 14 . Moreover, any fluorescence emissions 77 and 79 , corresponding respectively to light rays 76 and 78 will also be blocked by filter 18 , as they must be longer in wavelength than the bandpass of the filter region of filter 14 that they pass through, and they fall on filter regions of filter 18 that are formed by filter regions that have shorter wavelength bandpass characteristics.
  • light ray 80 has a wavelength corresponding to filter region 28 , and thus more energy than light passed by filter region 36 .
  • the sample will fluoresce with a lower energy and correspondingly longer wavelength light ray 81 that will pass through filter region 36 of filter 18 .
  • highest energy light ray 82 which passes through filter region 26 and the sample may emit a low energy photon 83 , which passes through filter region 35 and falls on the CCD detector.
  • line 56 in the case where filter 14 is identical to filter 18 , is a simple diagonal line.
  • the layout of the various bandpass filter regions varies rather considerably. Accordingly, it is necessary to accommodate such variations if one cannot go to the trouble of trying to match identical filters very carefully.
  • Such variations may cause line 56 to shift to the position illustrated by reference number 56 a in FIG. 6. Such variation occurs because the distance of oval which the series of spectral filters is dispersed is greater in filter 18 as compared to filter 14 .
  • filters 14 and 18 are made by depositing stripes of material which form bandpass filters on a substrate.
  • maximizing the thinness of instrument 10 will also maximize performance. More precisely, improved performance can be obtained by minimizing the distance between the active filter layer of filters 14 and 18 as well as minimizing the distance between the active layer of filter 18 and the sensitive face of detector 20 . Thus, exceedingly thin substrates may be used to optimize the performance of the instrument.
  • FIG. 7 Yet another approach is illustrated in FIG. 7.
  • FIG. 7 the convention of labeling parts with identical or analogous functions with numbers which vary by multiples of 100 has been followed.
  • the inventive spectrofluorometer 110 is excited by excitation light 138 along path 112 .
  • Excitation light 138 first falls on filter 114 , causing it to pass through the active layer 115 of filter 114 on the far side of filter 114 .
  • Light 138 then passes through the sample in receiver or carrier 116 .
  • Light 138 then passes through the active layer 117 of filter 116 .
  • Active layers 115 and 117 are formed on the substrates of their respective filters. Such substrates may be glass, plastic or any other suitable material.
  • After passing through active layer 117 light 138 passes through the substrate of filter 116 and on to the sensitive face of detector 120 , from which it is sent to a computer or other suitable device for interpreting and displaying the output of the detector.
  • spectrofluorometer 220 is excited by excitation light 238 along path 212 .
  • Excitation light 238 first falls on filter 214 , causing it to pass through the active layer 215 of filter 214 on the far side of filter 214 .
  • Light 238 then passes through the sample in receiver or carrier 216 .
  • Light 238 then passes through the active filter layer 217 , which is disposed and manufactured onto the output face of carrier or receiver 216 .
  • active filter layer 217 may be disposed on and manufactured onto the input face of detector 220 .
  • light 238 passes onto the sensitive face of detector 220 , from which it is sent to a computer or other suitable device for interpreting and displaying the output of the detector.
  • the distance between filtered light exiting the first active bandpass layer in the inventive system 220 , and the sensitive face of detector 220 is minimized in FIG. 8. Accordingly, light which is not traveling perpendicular to the faces of the filters, then, accordingly, is dispersed in itself, travels over a minimized path length and, accordingly, the dispersion is minimized, thus eliminating the need for the focusing optics, which are so important in prior art systems.
  • FIG. 9 a spectrofluorometer 310 having the feature of being able to block the excitation wavelength of the system is illustrated. This is desirable because the amplitude of the excitation wavelength will often spread and overload the detector receiving light from adjacent filter regions.
  • the instrument illustrated in FIG. 9, operates in the same manner as the instrument illustrated in FIG. 4, except for this additional feature.
  • the filter 314 has a filter 314 , a sample carrier 316 , a filter 318 , and a detector 320 .
  • the characteristics of all of these systems is the same as the instrument illustrated in FIG. 4.
  • it also has a spectral band reject filter 354 , which is aligned, filter region by filter region, to substantially identically opposite filter 314 .
  • filter region 323 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 324 .
  • filter region 325 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 326 .
  • Filter region 327 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 328 .
  • Filter region 329 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 330 .
  • Filter region 331 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 332 .
  • Filter region 333 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 334 .
  • FIG. 10 Another embodiment, shown in FIG. 10, is substantially identical to the instrument of FIG. 9, except that active filter layer 415 of spectrofluorometer 410 is deposited on the substrate of filter 414 on the side of filter 414 closer to the sample to be analyzed, and active filter layers 417 and 455 are deposited on the sensitive face of CCD 420 (on the side of filter 414 closer to the sample to be analyzed). This is done in order to minimize the lengths of paths of dispersion, and thus minimize dispersion and optimize the operation of the instrument. Active filter layer 455 is identical to filter 354 in FIG. 9.
  • Active filter layer 415 is made by advancing a mask along the substrate of filter 414 having the width of one of the regions illustrated in the figure, from one region to the next and applying the appropriate multilayer structure at each position to give the desired stripe of bandpass material the desired optical bandpass characteristic.
  • Active filter layer 417 is made by performing the same process, first applying to the sensitive face of CCD 420 the same series of different multilayer structures at their respective positions to give the corresponding stripes of filter layer 417 the desired optical bandpass characteristic. CCD 420 is then rotated in the plane of its sensitive face by 90 degrees.
  • Active filter layer 455 is made by advancing, along the rotated substrate of CCD 420 , a mask having the width of one of the regions illustrated in FIG. 10, from one region to the next and applying the appropriate multilayer structure at each position to give the desired stripe of band reject material the desired optical band reject characteristic. When the process is completed, the result is a filter layer 455 is the band reject analog of bandpass filter layer 415 .
  • layers 417 and 455 may be reversed by reversing their order of deposit.
  • the active filter layers may be deposited on the sample receiver or carrier to provide sample carriers that have filter patterns which may embody the operation of any of the systems described above.
  • sample carriers may be specialized to optimize the analysis of certain classes of analysis tasks, such as blood work, where it may be desirable to perform special filtering, to block, transmit or study certain portions of the spectrum.
  • One or more filter layers may be placed on either or both sides of the sample carrier.

Abstract

Spectrofluorometer employing a pair of linear variable spectral filters to produce a three dimensional data output is disclosed. A collimated white light source is used that first passes through a first linear variable spectral filter, then through a sample where fluorescence occurs, then the resultant light passes through a second linear variable spectral light filter that is oriented at ninety degrees from the first filter. The light is then detected by a CCD sensor for conversion into data. This arrangement provides a very simple, rugged and compact instrument that can be used almost anywhere, such as at the scene of a contamination accident.

Description

    CROSS-REFERENCE TO A RELATED APPLICATION
  • This application is a continuation of commonly owned application Ser. No. 09/678,709, the disclosure of which is hereby incorporated herein by reference thereto; which is a continuation-in-part of application Ser. No. 09/443,392, filed Nov. 19, 1999.[0001]
  • BACKGROUND OF THE INVENTION
  • Fluorescence instrumentation has been used for many years to identify unknown materials. Generally, the principle involved is that a material excited with light of a particular wavelength will emit light energy in the form of an emission spectrum whose amplitude profile, over the range of wavelengths emitted, constitutes a “fingerprint” which can give the identity and nature of the unknown material. [0002]
  • In the most demanding applications, a sample is excited with light of a single wavelength and the fluorescence emission spectrum is recorded. The wavelength of the excitation source is then advanced incrementally along the range of excitation wavelengths of interest, and the process repeated to record the fluorescence emission spectrum at the incremented wavelength. The process is continued until the entire range of excitation wavelengths of interest has been covered by the instrument. The result is a highly accurate, so-called three-dimensional fluorescence emission spectrum, showing excitation wavelengths, corresponding emission wavelengths and their amplitudes. Such instruments are of particular interest in scientific research where subtle variations in the characteristics of the spectrum may contain useful information to understand the effects of relatively subtle changes in the system. Typically, instruments of this sort have resolutions on the order of between 0.1 to 0.5 nm. [0003]
  • However, many applications have far less demanding requirements. For example, if one is merely interested in identifying the identity of a particular sample of material, far less resolution will suffice. Accordingly, a class of instruments having resolutions on the order of five to ten nanometers have seen widespread application in industry. Typical applications include the identification of samples of such material as blood, oil, pollutants and the like. Such instruments differ from other fluorescence instruments in that they are designed to perform measurements much more quickly, by measuring the fluorescence of a material over a range of wavelengths simultaneously. [0004]
  • Such a prior art system is illustrated in FIG. 1. Measurement of the fluorescence spectrum is achieved by having a system which comprises an excitation spectrograph [0005] 1 which is used to excite a sample 2, typically contained in an elongated cuvette 3. The elongated cuvette 3 is excited by an elongated image of a spectrum extending from a low wavelength to a high wavelength.
  • This results in fluorescence emission by [0006] sample 2 in cuvette 3. The emission is received and collimated by a collimating concave mirror 4, which reflects the fluorescence emission to focusing concave mirror 5, which, in turn, focuses the emitted fluorescence light at a slit 6, through which the light which comprises the fluorescence emission passes to fall on the planar mirror 7. Planar mirror 7 reflects the light toward a spectrograph 8 formed by a concave aberration-corrected diffraction grating. Spectrograph 8 disperses a spectrum on a CCD detector 9 which in a single row of pixels can produce the complete emission spectrum of the excited material.
  • In a typical instrument of this type, a xenon source is imaged as a bright line placed over a cuvette in a vertical line. Thus, the full spectrum will excite any homogeneous sample placed in the sample compartment of the cuvette. The resulting fluorescence emission is dispersed orthogonally over the active area of a rectangular CCD, or charge-coupled device, which is, essentially, a two-dimensional array of light detectors. The horizontal axis of the CCD records the emission spectra at different excitation wavelengths along the vertical axis, and gives the intensity for each wavelength. Thus, this instrument will produce, for each wavelength in the range of excitation wavelengths, the spectrum of emitted wavelengths. For example, if the system has a resolution of 5 nm, and covers a range of 100 nm, one could view the output as twenty different spectra. [0007]
  • The ability to complete a reading of the emission spectrum simultaneously opens up many possibilities for enhanced performance functions. For example, a cuvette may be fed by a high pressure liquid chromatography column, allowing the facile real-time generation of fluorescence emission spectra of the various materials in a sample being analyzed by the chromatography column. [0008]
  • While this system has many advantages over the prior art systems which measured a fluorescence spectrum one wavelength at a time, it still had a number of deficiencies. First, the volume required for the system is relatively large and precludes use of the system in a compact system. Moreover, the system comprises numerous expensive parts, and costs may be prohibitive for many applications. In addition, assembly of the system is unduly expensive requiring careful alignment of parts to ensure proper operation of the system. Similarly, the system is not as rugged as other systems, and is liable to become misaligned during use on account of shock and vibration. Finally, the system is limited to producing a fluorescence spectrum. [0009]
  • SUMMARY OF THE INVENTION
  • The invention, as claimed, is intended to provide a remedy. It solves the problems of large size, lack of ruggedness and cost by providing a simple instrument that can be implemented in a compact design. In accordance with the present invention, an excitation light source provides optical radiation over a range of wavelengths or spectra for illuminating a sample. The inventive instrument performs fluoresence analysis of samples, and comprises a light source emitting light into an illumination light path, and a first spectral filter in the illumination light path for transmitting light within a selected wavelength range. This defines a sample illumination light path. A second spectral filter is spaced from the first spectral filter forming a sample receiving space therebetween. [0010]
  • The illumination light path passes through the first spectral filter. The sample receiver and the second spectral filter lie in the light path, and the second spectral filter is displaced angularly relative to the first spectral filter. A sensing element in the resultant light path measures absorption spectra and fluorescence light. The first spectral filter and the second spectral filter have a characteristic which varies along an axis thereof. In accordance with the preferred embodiment of the invention, the variable characteristic is a variable bandpass wavelength in various filter regions of the spectral filter. Also in accordance with the preferred embodiment, the second spectral filter is angularly displaced at a substantially othogonal angle. [0011]
  • The above described embodiment of the invention has the advantage of providing along a diagonal region of the CCD the absorption spectrum of the material sample under analysis. [0012]
  • In accordance with an alternative embodiment of the invention, a third spectral filter in the resultant light path is oriented in a direction, and position in a position which are substantially the same as the direction and position of the first spectral filter. This third filter serves the function of a blocking filter thereby preventing excitation light energy that has passed through a sample receiver from passing to the sensing element or CCD array.[0013]
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • One way of carrying out the invention is described in detail below with reference to the drawings which illustrate one or more specific embodiments of the invention and in which like reference characters represent like elements: [0014]
  • FIG. 1 is a schematic view of a typical prior art apparatus; [0015]
  • FIG. 2 is a schematic view of the present invention showing the compactness of the components; [0016]
  • FIG. 3 is an exploded schematic view of the apparatus of the present invention showing a pair of linear variable spectral filter and a CCD type of sensing element; [0017]
  • FIG. 4 is an exploded view similar to FIG. 3 including a cartridge containing a sample to be tested; [0018]
  • FIG. 5 is a schematic view of a CCD sensing element, as is employed in the embodiments illustrated in FIGS. 4 and 5; [0019]
  • FIG. 6 is a schematic view of a CCD sensing element, as is employed in the embodiments illustrated in FIGS. 4 and 5, illustrating the absorption spectrum position when filter elements are not matched in the system of the present invention; [0020]
  • FIG. 7 is a perspective view of an alternative embodiment of the present invention; [0021]
  • FIG. 8 is a perspective view of an alternative embodiment of the inventive spectrofluorometer incorporating a further improvement; [0022]
  • FIG. 9 is a view similar to FIG. 4 of an alternative embodiment of the invention including an excitation light blocking filter; and [0023]
  • FIG. 10 is a perspective view of an embodiment of the invention similar to that illustrated in FIG. 9 and incorporating minimized light paths.[0024]
  • DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • Referring now to FIG. 2 and [0025] 3, major components of a spectrofluorometer 10 are shown. Optical radiation traveling along an excitation light path 12 passes into a linear variable spectral filter 14.
  • [0026] Spectral filter 14 is a device which has bandpass wavelength characteristics which vary along its length. More particularly, at the bottom of filter 14, one wavelength would be passed in the region defined by the dashed lines. In the next filter region above that filter region like having a different wavelength will be passed, perhaps a wavelength which is 5 nm longer. This sort of device is made by advancing a mask having the width of one of the regions illustrated in dashed lines in the figure, from one discrete position to another and applying a different multilayer structure at each position to give the corresponding stripe of bandpass material the desired optical bandpass characteristic.
  • The manufacture of such a filter is known in the art and forms no part of the present invention. Such filters may be purchased on the open market and are available from, for example, Reynard Corporation under their catalog No. 4610. Such a filter has a spectral range of 400 to 700 nm. It is relatively small and compact, being 60 mm long, 25 mm wide and 5 mm thick. A typical spectrum length would be 44 mm, with dispersion varying between 0.12 and 0.17 mm/nm. [0027]
  • The linear variable spectral filters sold by this corporation tend to vary in their characteristics, with a spectrum [0028] length varying form 37 to 51 mm. matching of the filters used in the embodiment of FIG. 2 is desirable. Alternatively, a computer reading the output of the system may calibrate the software against a known source.
  • A [0029] sample receiver 16 is located between the first spectral filter 14 and a second linear variable spectral filter 18. Sample receiver 16 is a vessel which defines a volume for receiving a sample which is to be analyzed. It may be a rectangular solid made of glass, plastic or any suitable material. It may also be as simple as a glass slide with a smear of the sample, or even a solid film of the sample material, such as tissue, paper from a paper mill whose operation is being monitored, and so forth.
  • Such a sample may be a solution derived from a material being tested, blood, the output of an HPLC liquid chromatography column, or the like. If the output of an HPLC column is being monitored, the [0030] receiver 16 may have a liquid input port and a drain, and the dimensions of the receiver would be such that capillary action insures the presence of sample material throughout the excited regions of receiver 16. A close-coupled discharge (CCD) sensing element 20 measures the relative position and intensity of light rays traveling along a resultant light path 12. See FIG. 3.
  • Sensing [0031] element 20 is preferably a CCD type of sensor although other types can be used depending upon the type of excitation light used and the sample to be tested. In FIGS. 3 and 5, detector 20 is shown as a 36 element matrix detector. The small number of elements or pixels is merely for the convenience of illustration and the illustration of the principles of the invention. In a real device, the number of detectors easily ranges into the hundreds of thousands of elements, and, depending upon the performances desired and the nature of the software reading out the signal from the detector, the number of elements in detector 20 may range into the millions of pixels.
  • In principle, even film can be used in place of [0032] detector 20. An absorption spectrum and lamp profile (without sample) is shown as diagonal line 56 in FIG. 5. In connection with the preferred embodiment of the invention, a suitable sensing element is the CCD sold by Instruments SA on the Spectrum One. Each of these elements are described in detail below.
  • Referring back to FIG. 3, the borders defining the filter regions with different spectral characteristics in the first and second [0033] optical filters 14 and 18 are shown as dashed lines. First filter 14 is a linear variable spectral filter that changes its bandpass wavelength along the length or planar axis 15 of the filter. Wavelengths outside the desired transmission ranges are blocked by the respective filter regions.
  • In a preferred embodiment, the spectral range from 400 to 700 nm is oriented vertically, e.g., with shortest [0034] wavelength filter region 24 at the bottom, then longer wavelength filter region 26, still longer wavelength filter region 28, a filter region 30 which passes a range of wavelengths longer than those of filter region 28, a filter region 32 which passes a range of wavelengths longer than those of filter region 30, and the longest wavelength bandpass filter region than 34 at the top. While the invention has been implemented with a spectral filter having the aforementioned wavelength characteristics, other visible and non-visible bandpass characteristics can be used depending on the nature and characteristics of the sample to be tested.
  • The second [0035] optical filter 18 is substantially the same as the first optical filter 14 except that it is oriented in such a manner that its gradations are not in line with those of first filter 14. The strips defining the bandpass filter regions on filter 18 are preferably at ninety degrees to those of filter 14. The advantages of this relationship will now be described in connection with the operation of the inventive system.
  • A [0036] light source 36 which may comprise a xenon lamp whose output is collimated by a lens or reflector, or any other suitable optical components produces an excitation white light ray bundle 38, sometimes referred to as illumination light, that travels along excitation light path 12 with a wide range of wavelengths striking the surface of filter 14. As white light ray bundle 38 passes through filter 14, selected wavelengths are passed by each filter region, such that a wavelength “gradient” from short to long wavelengths is produced. This is referred to herein as a sample excitation light 42.
  • As sample excitation light [0037] 42 passes through second filter 18, only those wavelengths of light that are not blocked pass completely through the filter 18. Since filter 18 is oriented at a right angle to filter 14, most of sample excitation light 42 is blocked. By way of example, λ1 passes through filter 14 and filter 18, while λ2 passes through filter 14, but is blocked by filter 18. In this manner a diagonal spectral line 56 is transmitted onto sensing element 20. The theoretical center of this line it illustrated in FIG. 5 by phantom line 56. This intrinsic relationship between the two linear variable spectral filters provides for simplicity of design, ruggedness and compact size of the inventive spectrofluorometer 10.
  • Referring now to FIG. 4, a [0038] sample receiver 16 is located between filter 14 and filter 18. Sample receiver 16 may be any of a number of conventional sample holding types or techniques. As sample excitation light passes through sample 44 some of the light energy is converted into fluorescence emissions. The physics of this conversion are well understood and generally involve the photon of excitation radiation raising the energy level of electrons in the excited atom to a higher energy level or shell. When the electron snaps back into its unexcited state, it emits a photon with an energy level lower that the exciting photon, thus resulting in the fluorescence having a wavelength longer than the excitation wavelength.
  • Some of the sample excitation light is “absorbed” by [0039] sample 44 and does not contribute to the emission. The net result is to increase the kinetic energy of the atoms of the sample, and thus raise the temperature of the sample.
  • A resultant [0040] light ray bundle 50, exiting sample receiver 16, comprises light rays which have exited filter 14 and fluoresence emissions from molecules that have been excited by light rays which have exited filter 14. Resultant light ray bundle 50 then passes into filter 18 where a selected wavelengths of both spectral light and fluorescent light are selectively blocked along the spectral gradient. The portions of light ray bundle 50 passing through to sensing element 20 constitutes the absorption spectrum 52 of the material being analyzed and appears along imaginary line 56 in FIG. 5. This can be used to identify sample 44.
  • As may be understood with reference to FIG. 4, filters [0041] 14 and 18 are substantially identical, but are positioned with their bandpass filter strip filter regions 24-34 and 35-44 oriented at right angles to each other. In accordance with the preferred embodiment of the invention, filter region 24 has the same bandpass characteristic as filter region 34. In accordance with the preferred embodiment of the invention, filter region 26 has the same bandpass characteristic as filter region 42. Filter region 28 has the same bandpass characteristic as filter region 40. Filter region 30 has the same bandpass characteristic as filter region 37. Filter region 32 has the same bandpass characteristic as filter region 36. Filter region 34 has the same bandpass characteristic as filter region 35.
  • Thus, the [0042] CCD elements 70, lying along line 56 in FIG. 5, are the only elements that will be illuminated by the white light ray bundle 38 coming from the excitation source. Moreover, because the fluorescence spectrum constitutes only wavelengths longer than the excitation wavelength, they will be blocked from reaching elements 70 by filter 18. Thus, only the absorption spectrum can be seen along imaginary line 56 to provide a first identification of the sample.
  • Likewise, because the fluorescence spectrum constitutes only wavelengths longer than the excitation wavelength, these longer wavelengths will be passed by [0043] filter 18 to those elements 58 of the CCD which lie below line 56 in FIG. 5. Thus, the elements 58 of the CCD which lie below line 56 in FIG. 5 produce the fluorescence emission spectra of the sample under analysis. The resultant fluorescence emission is used to identify sample 44.
  • Referring back to FIG. 4, the operation of the inventive system may be better understood. In particular, the output of the [0044] xenon lamp 36 constituting a broadband emission which is collimated into white light ray bundle 38 is caused to fall on filter 14, which outputs a plurality of stripes of light energy at different wavelengths. Because filters 14 and 18 are very thin, as is sample container 16, the output of filter 14 is effectively “imaged” on the sample in sample receiver 16. The output of sample container 16 is likewise effectively “imaged” on filter 18. Finally, in turn, the output of filter 18 is effectively “imaged” on the surface of CCD elements 58. The system works because all of the above thin elements are in contact with each other and CCD 20 to form the sandwich illustrated in FIG. 2.
  • As noted above, [0045] light ray 72, which is one of the light rays in white light bundle 38, because it is in the bandpass range of filter region 34 on filter 14, and, naturally, in the bandpass of optically identical filter region 35, will pass through both filters and fall on CCD 20, if it is not absorbed by the sample. The same is true for light ray 74, which is in the bandpass of filter regions 24 and 44.
  • Light rays [0046] 76 and 78 will, on the other hand, be blocked by filter 18, after being limited to the different bandpass of facing filter regions of filter 14. Moreover, any fluorescence emissions 77 and 79, corresponding respectively to light rays 76 and 78 will also be blocked by filter 18, as they must be longer in wavelength than the bandpass of the filter region of filter 14 that they pass through, and they fall on filter regions of filter 18 that are formed by filter regions that have shorter wavelength bandpass characteristics.
  • In contrast, [0047] light ray 80 has a wavelength corresponding to filter region 28, and thus more energy than light passed by filter region 36. Thus, it is physically possible that the sample will fluoresce with a lower energy and correspondingly longer wavelength light ray 81 that will pass through filter region 36 of filter 18. Likewise, highest energy light ray 82 which passes through filter region 26 and the sample may emit a low energy photon 83, which passes through filter region 35 and falls on the CCD detector.
  • Conversely, it is physically impossible that a sample will fluoresce with a higher energy and correspondingly shorter wavelength. Thus, a photon of [0048] light energy 84 passing through filter region 34 of filter 18 has the lowest energy in the system and the sample cannot emit a higher energy photon, and thus any light 85, whether transmitted or emitted by the sample will be blocked by filter region 38 which has a shorter bandpass wavelength than filter region 34. Thus, any such light will not reach the CCD detector.
  • Referring to FIG. 6, it can be seen that [0049] line 56, in the case where filter 14 is identical to filter 18, is a simple diagonal line. However, due to the nature of the manufacturing process use to produce filters 14 and 18, the layout of the various bandpass filter regions varies rather considerably. Accordingly, it is necessary to accommodate such variations if one cannot go to the trouble of trying to match identical filters very carefully.
  • Such variations may cause [0050] line 56 to shift to the position illustrated by reference number 56 a in FIG. 6. Such variation occurs because the distance of oval which the series of spectral filters is dispersed is greater in filter 18 as compared to filter 14.
  • In the case of such variations, it is merely necessary to calibrate the software to the pattern on [0051] CCD 20. This can be done by determining the presence of the absorption spectrum and then mathematically adjusting the position of the fluorescence spectrum accordingly. This is done on the basis that the opposite ends of the absorption spectrum represent the horizontal and vertical limits of the fluorescence spectrum. Such determination can most easily be made without having a sample in the inventive fluorescence instrument 10.
  • As is alluded to above, filters [0052] 14 and 18 are made by depositing stripes of material which form bandpass filters on a substrate. As is also alluded to above, maximizing the thinness of instrument 10 will also maximize performance. More precisely, improved performance can be obtained by minimizing the distance between the active filter layer of filters 14 and 18 as well as minimizing the distance between the active layer of filter 18 and the sensitive face of detector 20. Thus, exceedingly thin substrates may be used to optimize the performance of the instrument.
  • Yet another approach is illustrated in FIG. 7. In FIG. 7 the convention of labeling parts with identical or analogous functions with numbers which vary by multiples of [0053] 100 has been followed.
  • In FIG. 7, the [0054] inventive spectrofluorometer 110 is excited by excitation light 138 along path 112. Excitation light 138 first falls on filter 114, causing it to pass through the active layer 115 of filter 114 on the far side of filter 114. Light 138 then passes through the sample in receiver or carrier 116. Light 138 then passes through the active layer 117 of filter 116. Active layers 115 and 117 are formed on the substrates of their respective filters. Such substrates may be glass, plastic or any other suitable material. After passing through active layer 117, light 138 passes through the substrate of filter 116 and on to the sensitive face of detector 120, from which it is sent to a computer or other suitable device for interpreting and displaying the output of the detector.
  • Yet another approach is shown in FIG. 8. Here spectrofluorometer [0055] 220 is excited by excitation light 238 along path 212. Excitation light 238 first falls on filter 214, causing it to pass through the active layer 215 of filter 214 on the far side of filter 214. Light 238 then passes through the sample in receiver or carrier 216. Light 238 then passes through the active filter layer 217, which is disposed and manufactured onto the output face of carrier or receiver 216. Alternatively, active filter layer 217 may be disposed on and manufactured onto the input face of detector 220. After passing through active layer 217, light 238 passes onto the sensitive face of detector 220, from which it is sent to a computer or other suitable device for interpreting and displaying the output of the detector.
  • As will the apparent from FIG. 8, the distance between filtered light exiting the first active bandpass layer in the [0056] inventive system 220, and the sensitive face of detector 220 is minimized in FIG. 8. Accordingly, light which is not traveling perpendicular to the faces of the filters, then, accordingly, is dispersed in itself, travels over a minimized path length and, accordingly, the dispersion is minimized, thus eliminating the need for the focusing optics, which are so important in prior art systems.
  • Referring to FIG. 9, a [0057] spectrofluorometer 310 having the feature of being able to block the excitation wavelength of the system is illustrated. This is desirable because the amplitude of the excitation wavelength will often spread and overload the detector receiving light from adjacent filter regions. The instrument illustrated in FIG. 9, operates in the same manner as the instrument illustrated in FIG. 4, except for this additional feature.
  • In particular, it has a [0058] filter 314, a sample carrier 316, a filter 318, and a detector 320. The characteristics of all of these systems is the same as the instrument illustrated in FIG. 4. However, it also has a spectral band reject filter 354, which is aligned, filter region by filter region, to substantially identically opposite filter 314.
  • More particularly, in accordance with the preferred embodiment of the invention, [0059] filter region 323 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 324. In accordance with the preferred embodiment of the invention, filter region 325 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 326. Filter region 327 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 328. Filter region 329 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 330. Filter region 331 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 332. Filter region 333 has a band reject characteristic with the same wavelength range as the wavelength range of the bandpass characteristic of filter region 334. The blocking of excitation wavelengths is thus assured and the detection of low amplitude fluorescence signals is enhanced.
  • Another embodiment, shown in FIG. 10, is substantially identical to the instrument of FIG. 9, except that [0060] active filter layer 415 of spectrofluorometer 410 is deposited on the substrate of filter 414 on the side of filter 414 closer to the sample to be analyzed, and active filter layers 417 and 455 are deposited on the sensitive face of CCD 420 (on the side of filter 414 closer to the sample to be analyzed). This is done in order to minimize the lengths of paths of dispersion, and thus minimize dispersion and optimize the operation of the instrument. Active filter layer 455 is identical to filter 354 in FIG. 9.
  • [0061] Active filter layer 415 is made by advancing a mask along the substrate of filter 414 having the width of one of the regions illustrated in the figure, from one region to the next and applying the appropriate multilayer structure at each position to give the desired stripe of bandpass material the desired optical bandpass characteristic. Active filter layer 417 is made by performing the same process, first applying to the sensitive face of CCD 420 the same series of different multilayer structures at their respective positions to give the corresponding stripes of filter layer 417 the desired optical bandpass characteristic. CCD 420 is then rotated in the plane of its sensitive face by 90 degrees.
  • [0062] Active filter layer 455 is made by advancing, along the rotated substrate of CCD 420, a mask having the width of one of the regions illustrated in FIG. 10, from one region to the next and applying the appropriate multilayer structure at each position to give the desired stripe of band reject material the desired optical band reject characteristic. When the process is completed, the result is a filter layer 455 is the band reject analog of bandpass filter layer 415.
  • In accordance with the present invention, it is may be desirable, in order to accommodate the insertion of different sample receivers or [0063] carriers 416, to vary the distance between filter layers 415 and 417. This may be achieved by mounting filter 414 on a horizontally moveable table 491 or other mechanism. This enables movement in the directions indicated by arrow 492.
  • The positions of [0064] layers 417 and 455 may be reversed by reversing their order of deposit. Likewise, the active filter layers may be deposited on the sample receiver or carrier to provide sample carriers that have filter patterns which may embody the operation of any of the systems described above. Such sample carriers may be specialized to optimize the analysis of certain classes of analysis tasks, such as blood work, where it may be desirable to perform special filtering, to block, transmit or study certain portions of the spectrum. One or more filter layers may be placed on either or both sides of the sample carrier.
  • While an illustrative embodiment of the invention has been described, it is, of course, understood that various modifications of the invention may be made by those of ordinary skill in the art without departing from the spirit and scope of the invention which is limited and defined only by the appended claims. [0065]

Claims (23)

1. An instrument for measuring the wavelength characteristics of light output from a material in response to a light input, comprising:
(a) a light source for producing input light;
(b) a first filter, said first filter defining a first active area, said first filter being positioned to receive said input light from said source, said first filter having a characteristic that varies from position to position along said first active area, said first filter transmitting a portion of said input light through said first filter as first filtered light;
(c) a second filter, said second filter defining a second active area, said second filter being positioned to receive said first filtered light from said first filter, said second filter being positioned in facing spaced relationship to said first filter to define a space for the placement of a sample to be analyzed, said second filter having a characteristic that varies from position to position along said second active area, said second filter transmitting a portion of said input first filtered light through said second filter as second filtered light, at least some of the facing portions of said second filter facing said first filter having a transmissive characteristic different from that of the facing portion of said first filter; and
(d) a detector for detecting said second filtered light.
2. An instrument as in
claim 1
, wherein said first and second filters have a bandpass characteristic which varies from position to position to allow the measurement of an emission spectrum.
3. An instrument as in
claim 2
, wherein said first and second filters each comprise a series of strips with different bandpass characteristics, and are angularly positioned with respect to each other.
4. An instrument as in
claim 3
, wherein said detector is a two dimensional array.
5. An instrument as in
claim 4
, wherein said first and second filters are positioned at substantially a right angle with respect to each other.
6. An instrument as in
claim 4
, wherein said first and second filters are positioned with their active surfaces facing toward each other.
7. An instrument as in
claim 6
, wherein said first and second filters are positionable at a variable distance with respect to each other.
8. An instrument as in
claim 4
, wherein said first and second filters are positionable at a variable distance with respect to each other.
9. An instrument as in
claim 4
, further comprising a third filter, said third filter defining a third active area, said third filter being positioned to receive said second filtered light from said second filter, said third filter having a characteristic that is substantially a band reject analog of the bandpass characteristic of said first filter, said third filter blocking said first filtered light and transmitting at least a portion of said input second filtered light through said third filter as third filtered light, said third filtered light passing on to said detector, whereby the excitation wavelengths do not overload the detector and hinder the detection of fluorescence emissions.
10. An instrument as in
claim 9
, wherein said first and second filters are positioned at substantially a right angle with respect to each other.
11. An instrument as in
claim 9
, wherein said first and second filters are positioned with their active surfaces facing toward each other
12. An instrument as in
claim 2
, wherein said first filter has first portions with the same bandpass characteristics as a facing portion of said second filter to allow the measurement of an absorption spectrum and wherein said detector measures fluorescence spectra.
13. An instrument as in
claim 12
, wherein said first and second filters each comprise a series of strips with different bandpass characteristics.
14. An instrument as in
claim 13
, wherein said detector is a two dimensional array.
15. An instrument for spectro-fluoresence analysis of samples, said instrument comprising:
a light source emitting light along an illumination light path;
a first spectral filter in said illumination light path for receiving the output of said light source, and transmitting light within a selected wavelength range;
a second spectral filter, said second spectral filter spaced from said first spectral filter forming a sample receiver therebetween, said illumination light path passing through said first spectral filter, said sample receiver and said second spectral filter, said second spectral filter being displaced angularly relative to said first spectral filter; and
a sensing element in said resultant light path for measuring absorption spectra and fluorescence light.
16. An instrument according to
claim 15
, wherein said first spectral filter and said second spectral filter include a variable characteristic along an axis thereof.
17. An instrument according to
claim 16
, wherein said variable characteristic includes different bandpass regions across said axis.
18. An instrument according to
claim 17
, wherein said second spectral filter is angularly displaced at a substantially othogonal angle.
19. An instrument according to
claim 15
, wherein a third spectral filter in said resultant light path is characterized in that it is oriented substantially the same as said first spectral filter and is a blocking filter with a wavelength characteristic that prevents illumination light that has passed through said sample receiver from passing through said third spectral filter.
20. An instrument according to
claim 15
, wherein said sensing element is a CCD.
21. An instrument for spectral analysis of samples, said instrument comprising:
a collimated white light source;
a linear vertically variable spectral filter;
a linear horizontally variable spectral filter;
a sensing element; and
a support structure for holding said collimated white light source, said linear vertically variable spectral filter, said linear horizontally variable spectral filter, and said sensing element along a light path, whereby, when said collimated white light source is energized, light will pass through portions of said linear vertically variable spectral filter then pass through said linear horizontally variable spectral filter and into said sensing element producing a profile of said collimated white light source.
22. A fluorescence imaging apparatus for measuring radiation emitted by a sample to be tested as a result of the fluorescing of a sample excited by illuminating radiation, said apparatus comprising:
an illumination source producing optical radiation;
a first optical filter having an active filter area for transmitting said optical radiation within a selected wavelength range;
a second optical filter spaced from said first optical filter forming a sample receiving volume therebetween, said second optical filter including a characteristic of blocking fluorescence light generated in said sample receiver in a range of selected wavelengths at a portion of its active filtering area; and
a sensing element for receiving spectral radiation and fluorescing radiation.
23. An instrument for measuring the wavelength characteristics of light output from a material in response to a light input, comprising:
(a) a light source for producing input light;
(b) a first active filter area, said first active filter area defining a first active area, said first active filter area being positioned to receive said input light from said source, said first active filter area having a characteristic that varies from position to position along said first active area, said first active filter area transmitting a portion of said input light through said first active filter area as first active filter areaed light;
(c) a second active filter area, said second active filter area defining a second active area, said second active filter area being positioned to receive said first filtered light from said first active filter area, said second active filter area being positioned in facing spaced relationship to said first active filter area to define a space for the placement of a sample to be analyzed, said second active filter area having a characteristic that varies from position to position along said second active area, said second active filter area transmitting a portion of said input first filtered light through said second active filter area as second filtered light, at least some of the facing portions of said second active filter area facing said first active filter area having a transmissive characteristic different from that of the facing portion of said first active filter area; and
(d) a detector having an active face for detecting said second filtered light, said second active filter area being disposed on and secured to said active face of said CCD or on a sample carrier disposed between said first active filter area and said second active filter area.
US09/813,325 1999-11-19 2001-03-19 Compact spectrofluorometer Expired - Lifetime US6441892B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US09/813,325 US6441892B2 (en) 1999-11-19 2001-03-19 Compact spectrofluorometer

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US09/443,392 US6323944B1 (en) 1999-11-19 1999-11-19 Compact spectrofluorometer
US67870900A 2000-10-04 2000-10-04
US09/813,325 US6441892B2 (en) 1999-11-19 2001-03-19 Compact spectrofluorometer

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
US67870900A Continuation 1999-11-19 2000-10-04

Publications (2)

Publication Number Publication Date
US20010028458A1 true US20010028458A1 (en) 2001-10-11
US6441892B2 US6441892B2 (en) 2002-08-27

Family

ID=27033512

Family Applications (1)

Application Number Title Priority Date Filing Date
US09/813,325 Expired - Lifetime US6441892B2 (en) 1999-11-19 2001-03-19 Compact spectrofluorometer

Country Status (6)

Country Link
US (1) US6441892B2 (en)
EP (1) EP1232387A4 (en)
JP (1) JP2003515129A (en)
CN (1) CN1409818A (en)
CA (1) CA2392228A1 (en)
WO (1) WO2001036948A1 (en)

Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050046839A1 (en) * 2003-07-16 2005-03-03 Agence Spatiale Europeenne Miniature optical spectrometer
US20070002325A1 (en) * 2005-02-17 2007-01-04 Jimpei Tabata Fluorescence measurement apparatus
US20070188760A1 (en) * 2006-02-15 2007-08-16 Ahmed Bouzid Fluorescence filtering system and method for molecular imaging
US20080239070A1 (en) * 2006-12-22 2008-10-02 Novadaq Technologies Inc. Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US20090080194A1 (en) * 2006-02-15 2009-03-26 Li-Cor, Inc. Fluorescence filtering system and method for molecular imaging
US20090095885A1 (en) * 2007-10-11 2009-04-16 Hager Harold E System and methods for detecting semiconductor-based photodiodes
US20150103229A1 (en) * 2013-02-13 2015-04-16 Panasonic Intellectual Property Management Co., Ltd. Multispectral imaging device and multispectral imaging method
KR20160092085A (en) 2015-01-26 2016-08-04 이화여자대학교 산학협력단 Proactive portable algae detecting method and apparatus
KR20160131133A (en) 2015-05-06 2016-11-16 이화여자대학교 산학협력단 Control method for portable algae detecting apparatus and portable algae detecting apparatus
KR101683379B1 (en) 2015-05-29 2016-12-06 이화여자대학교 산학협력단 Portable algae detecting apparatus
US9541750B2 (en) 2014-06-23 2017-01-10 Li-Cor, Inc. Telecentric, wide-field fluorescence scanning systems and methods
KR20170015959A (en) 2017-02-01 2017-02-10 이화여자대학교 산학협력단 Proactive portable algae detecting apparatus
US9642532B2 (en) 2008-03-18 2017-05-09 Novadaq Technologies Inc. Imaging system for combined full-color reflectance and near-infrared imaging
CN107085291A (en) * 2016-02-15 2017-08-22 徕卡仪器(新加坡)有限公司 For the microscopical illumination filter system of multispectral fluorescence of performing the operation and its illumination wave filter
EP3100078A4 (en) * 2014-01-31 2017-09-06 JDS Uniphase Corporation An optical filter and spectrometer
US9814378B2 (en) 2011-03-08 2017-11-14 Novadaq Technologies Inc. Full spectrum LED illuminator having a mechanical enclosure and heatsink
US9945790B2 (en) 2015-08-05 2018-04-17 Viavi Solutions Inc. In-situ spectral process monitoring
US10048127B2 (en) 2015-08-05 2018-08-14 Viavi Solutions Inc. Optical filter and spectrometer
US10869645B2 (en) 2016-06-14 2020-12-22 Stryker European Operations Limited Methods and systems for adaptive imaging for low light signal enhancement in medical visualization
USD916294S1 (en) 2016-04-28 2021-04-13 Stryker European Operations Limited Illumination and imaging device
US10980420B2 (en) 2016-01-26 2021-04-20 Stryker European Operations Limited Configurable platform
US10992848B2 (en) 2017-02-10 2021-04-27 Novadaq Technologies ULC Open-field handheld fluorescence imaging systems and methods
US11930278B2 (en) 2015-11-13 2024-03-12 Stryker Corporation Systems and methods for illumination and imaging of a target

Families Citing this family (79)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE454845T1 (en) 2000-10-30 2010-01-15 Gen Hospital Corp OPTICAL SYSTEMS FOR TISSUE ANALYSIS
US9295391B1 (en) 2000-11-10 2016-03-29 The General Hospital Corporation Spectrally encoded miniature endoscopic imaging probe
DE10297689B4 (en) 2001-05-01 2007-10-18 The General Hospital Corp., Boston Method and device for the determination of atherosclerotic coating by measurement of optical tissue properties
US7355716B2 (en) 2002-01-24 2008-04-08 The General Hospital Corporation Apparatus and method for ranging and noise reduction of low coherence interferometry LCI and optical coherence tomography OCT signals by parallel detection of spectral bands
US7643153B2 (en) 2003-01-24 2010-01-05 The General Hospital Corporation Apparatus and method for ranging and noise reduction of low coherence interferometry LCI and optical coherence tomography OCT signals by parallel detection of spectral bands
EP1596716B1 (en) 2003-01-24 2014-04-30 The General Hospital Corporation System and method for identifying tissue using low-coherence interferometry
EP1611470B1 (en) 2003-03-31 2015-10-14 The General Hospital Corporation Speckle reduction in optical coherence tomography by path length encoded angular compounding
EP2008579B1 (en) 2003-06-06 2016-11-09 The General Hospital Corporation Process and apparatus for a wavelength tuned light source
JP5567246B2 (en) 2003-10-27 2014-08-06 ザ ジェネラル ホスピタル コーポレイション Method and apparatus for performing optical imaging using frequency domain interferometry
EP1754016B1 (en) 2004-05-29 2016-05-18 The General Hospital Corporation Process, system and software arrangement for a chromatic dispersion compensation using reflective layers in optical coherence tomography (oct) imaging
WO2006014392A1 (en) 2004-07-02 2006-02-09 The General Hospital Corporation Endoscopic imaging probe comprising dual clad fibre
US7532326B2 (en) * 2004-07-07 2009-05-12 Corcoran Timothy C Multiple-label fluorescence imaging using excitation-emission matrices
EP1782020B1 (en) 2004-08-06 2012-10-03 The General Hospital Corporation Process, system and software arrangement for determining at least one location in a sample using an optical coherence tomography
ATE538714T1 (en) 2004-08-24 2012-01-15 Gen Hospital Corp METHOD, SYSTEM AND SOFTWARE ARRANGEMENT FOR DETERMINING THE ELASTIC MODULE
KR20070058523A (en) 2004-08-24 2007-06-08 더 제너럴 하스피탈 코포레이션 Method and apparatus for imaging of vessel segments
EP1787105A2 (en) 2004-09-10 2007-05-23 The General Hospital Corporation System and method for optical coherence imaging
JP4997112B2 (en) 2004-09-29 2012-08-08 ザ ジェネラル ホスピタル コーポレイション Apparatus for transmitting at least one electromagnetic radiation and method of manufacturing the same
EP2278265A3 (en) 2004-11-24 2011-06-29 The General Hospital Corporation Common-Path Interferometer for Endoscopic OCT
EP1816949A1 (en) 2004-11-29 2007-08-15 The General Hospital Corporation Arrangements, devices, endoscopes, catheters and methods for performing optical imaging by simultaneously illuminating and detecting multiple points on a sample
ES2337497T3 (en) 2005-04-28 2010-04-26 The General Hospital Corporation EVALUATION OF CHARACTERISTICS OF THE IMAGE OF AN ANATOMICAL STRUCTURE IN IMAGES OF TOMOGRAPHY OF OPTICAL COHERENCE.
EP1889037A2 (en) 2005-06-01 2008-02-20 The General Hospital Corporation Apparatus, method and system for performing phase-resolved optical frequency domain imaging
CN101238347B (en) 2005-08-09 2011-05-25 通用医疗公司 Apparatus, methods and storage medium for performing polarization-based quadrature demodulation in optical coherence tomography
CN101304683B (en) 2005-09-29 2012-12-12 通用医疗公司 Method and apparatus for method for viewing and analyzing of one or more biological samples with progressively increasing resolutions
WO2007047690A1 (en) 2005-10-14 2007-04-26 The General Hospital Corporation Spectral- and frequency- encoded fluorescence imaging
EP1971848B1 (en) 2006-01-10 2019-12-04 The General Hospital Corporation Systems and methods for generating data based on one or more spectrally-encoded endoscopy techniques
US9087368B2 (en) 2006-01-19 2015-07-21 The General Hospital Corporation Methods and systems for optical imaging or epithelial luminal organs by beam scanning thereof
US8145018B2 (en) 2006-01-19 2012-03-27 The General Hospital Corporation Apparatus for obtaining information for a structure using spectrally-encoded endoscopy techniques and methods for producing one or more optical arrangements
WO2007149602A2 (en) 2006-02-01 2007-12-27 The General Hospital Corporation Methods and systems for providing electromagnetic radiation to at least one portion of a sample using conformal laser therapy procedures
US9186066B2 (en) 2006-02-01 2015-11-17 The General Hospital Corporation Apparatus for applying a plurality of electro-magnetic radiations to a sample
JP5519152B2 (en) 2006-02-08 2014-06-11 ザ ジェネラル ホスピタル コーポレイション Device for acquiring information about anatomical samples using optical microscopy
US7982879B2 (en) 2006-02-24 2011-07-19 The General Hospital Corporation Methods and systems for performing angle-resolved fourier-domain optical coherence tomography
JP2009536740A (en) 2006-05-10 2009-10-15 ザ ジェネラル ホスピタル コーポレイション Process, configuration and system for providing frequency domain imaging of samples
JP2010501877A (en) 2006-08-25 2010-01-21 ザ ジェネラル ホスピタル コーポレイション Apparatus and method for improving optical coherence tomography imaging capabilities using volumetric filtering techniques
US8838213B2 (en) 2006-10-19 2014-09-16 The General Hospital Corporation Apparatus and method for obtaining and providing imaging information associated with at least one portion of a sample, and effecting such portion(s)
US8155730B2 (en) 2006-10-24 2012-04-10 The Research Foundation Of State University Of New York Composition, method, system, and kit for optical electrophysiology
EP2662674A3 (en) 2007-01-19 2014-06-25 The General Hospital Corporation Rotating disk reflection for fast wavelength scanning of dispersed broadbend light
WO2008118781A2 (en) 2007-03-23 2008-10-02 The General Hospital Corporation Methods, arrangements and apparatus for utilizing a wavelength-swept laser using angular scanning and dispersion procedures
US10534129B2 (en) 2007-03-30 2020-01-14 The General Hospital Corporation System and method providing intracoronary laser speckle imaging for the detection of vulnerable plaque
US8045177B2 (en) 2007-04-17 2011-10-25 The General Hospital Corporation Apparatus and methods for measuring vibrations using spectrally-encoded endoscopy
WO2009018456A2 (en) 2007-07-31 2009-02-05 The General Hospital Corporation Systems and methods for providing beam scan patterns for high speed doppler optical frequency domain imaging
US7933021B2 (en) 2007-10-30 2011-04-26 The General Hospital Corporation System and method for cladding mode detection
US7898656B2 (en) * 2008-04-30 2011-03-01 The General Hospital Corporation Apparatus and method for cross axis parallel spectroscopy
EP2274572A4 (en) 2008-05-07 2013-08-28 Gen Hospital Corp System, method and computer-accessible medium for tracking vessel motion during three-dimensional coronary artery microscopy
JP5795531B2 (en) 2008-06-20 2015-10-14 ザ ジェネラル ホスピタル コーポレイション Fused fiber optic coupler structure and method of using the same
US9254089B2 (en) 2008-07-14 2016-02-09 The General Hospital Corporation Apparatus and methods for facilitating at least partial overlap of dispersed ration on at least one sample
US8440952B2 (en) * 2008-11-18 2013-05-14 The Regents Of The University Of California Methods for optical amplified imaging using a two-dimensional spectral brush
JP5731394B2 (en) 2008-12-10 2015-06-10 ザ ジェネラル ホスピタル コーポレイション System, apparatus and method for extending imaging depth range of optical coherence tomography through optical subsampling
WO2010090837A2 (en) 2009-01-20 2010-08-12 The General Hospital Corporation Endoscopic biopsy apparatus, system and method
US8097864B2 (en) 2009-01-26 2012-01-17 The General Hospital Corporation System, method and computer-accessible medium for providing wide-field superresolution microscopy
CA2749670A1 (en) 2009-02-04 2010-08-12 The General Hospital Corporation Apparatus and method for utilization of a high-speed optical wavelength tuning source
CN102469943A (en) 2009-07-14 2012-05-23 通用医疗公司 Apparatus, systems and methods for measuring flow and pressure within a vessel
HUE052561T2 (en) 2010-03-05 2021-05-28 Massachusetts Gen Hospital Apparatus for providing electro-magnetic radiation to a sample
US9069130B2 (en) 2010-05-03 2015-06-30 The General Hospital Corporation Apparatus, method and system for generating optical radiation from biological gain media
US9557154B2 (en) 2010-05-25 2017-01-31 The General Hospital Corporation Systems, devices, methods, apparatus and computer-accessible media for providing optical imaging of structures and compositions
JP5778762B2 (en) 2010-05-25 2015-09-16 ザ ジェネラル ホスピタル コーポレイション Apparatus and method for spectral analysis of optical coherence tomography images
EP2575591A4 (en) 2010-06-03 2017-09-13 The General Hospital Corporation Apparatus and method for devices for imaging structures in or at one or more luminal organs
EP2632324A4 (en) 2010-10-27 2015-04-22 Gen Hospital Corp Apparatus, systems and methods for measuring blood pressure within at least one vessel
US9330092B2 (en) 2011-07-19 2016-05-03 The General Hospital Corporation Systems, methods, apparatus and computer-accessible-medium for providing polarization-mode dispersion compensation in optical coherence tomography
US10241028B2 (en) 2011-08-25 2019-03-26 The General Hospital Corporation Methods, systems, arrangements and computer-accessible medium for providing micro-optical coherence tomography procedures
EP2769491A4 (en) 2011-10-18 2015-07-22 Gen Hospital Corp Apparatus and methods for producing and/or providing recirculating optical delay(s)
EP2833776A4 (en) 2012-03-30 2015-12-09 Gen Hospital Corp Imaging system, method and distal attachment for multidirectional field of view endoscopy
US8906320B1 (en) 2012-04-16 2014-12-09 Illumina, Inc. Biosensors for biological or chemical analysis and systems and methods for same
US11490797B2 (en) 2012-05-21 2022-11-08 The General Hospital Corporation Apparatus, device and method for capsule microscopy
WO2014031748A1 (en) 2012-08-22 2014-02-27 The General Hospital Corporation System, method, and computer-accessible medium for fabrication minature endoscope using soft lithography
JP6560126B2 (en) 2013-01-28 2019-08-14 ザ ジェネラル ホスピタル コーポレイション Apparatus and method for providing diffusion spectroscopy superimposed on optical frequency domain imaging
US10893806B2 (en) 2013-01-29 2021-01-19 The General Hospital Corporation Apparatus, systems and methods for providing information regarding the aortic valve
US11179028B2 (en) 2013-02-01 2021-11-23 The General Hospital Corporation Objective lens arrangement for confocal endomicroscopy
JP6378311B2 (en) 2013-03-15 2018-08-22 ザ ジェネラル ホスピタル コーポレイション Methods and systems for characterizing objects
US9784681B2 (en) 2013-05-13 2017-10-10 The General Hospital Corporation System and method for efficient detection of the phase and amplitude of a periodic modulation associated with self-interfering fluorescence
WO2015010133A1 (en) 2013-07-19 2015-01-22 The General Hospital Corporation Determining eye motion by imaging retina. with feedback
EP3692887B1 (en) 2013-07-19 2024-03-06 The General Hospital Corporation Imaging apparatus which utilizes multidirectional field of view endoscopy
EP3910282B1 (en) 2013-07-26 2024-01-17 The General Hospital Corporation Method of providing a laser radiation with a laser arrangement utilizing optical dispersion for applications in fourier-domain optical coherence tomography
EP4220137A1 (en) 2013-12-10 2023-08-02 Illumina, Inc. Biosensors for biological or chemical analysis and methods of manufacturing the same
US9733460B2 (en) 2014-01-08 2017-08-15 The General Hospital Corporation Method and apparatus for microscopic imaging
US10736494B2 (en) 2014-01-31 2020-08-11 The General Hospital Corporation System and method for facilitating manual and/or automatic volumetric imaging with real-time tension or force feedback using a tethered imaging device
WO2015153982A1 (en) 2014-04-04 2015-10-08 The General Hospital Corporation Apparatus and method for controlling propagation and/or transmission of electromagnetic radiation in flexible waveguide(s)
ES2907287T3 (en) 2014-07-25 2022-04-22 Massachusetts Gen Hospital Apparatus for imaging and in vivo diagnosis
US10861829B2 (en) 2017-12-26 2020-12-08 Illumina, Inc. Sensor system
CA3127027A1 (en) * 2019-01-17 2020-07-23 University Health Network Tissue phantoms

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2663801A (en) * 1951-10-08 1953-12-22 Slavin Morris Transmission fluorimeter
US4031398A (en) * 1976-03-23 1977-06-21 Research Corporation Video fluorometer
US4937457A (en) 1989-02-10 1990-06-26 Slm Instruments, Inc. Picosecond multi-harmonic fourier fluorometer
US5721613A (en) * 1994-03-21 1998-02-24 Hewlett Packard Company Fluorescence spectrometer
US5943129A (en) 1997-08-07 1999-08-24 Cambridge Research & Instrumentation Inc. Fluorescence imaging system
US6140653A (en) * 1998-03-27 2000-10-31 Vysis, Inc. Large-field fluorescence imaging apparatus

Cited By (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7019833B2 (en) * 2003-07-16 2006-03-28 Agence Spatiale Europeenne Miniature optical spectrometer
US20050046839A1 (en) * 2003-07-16 2005-03-03 Agence Spatiale Europeenne Miniature optical spectrometer
US20070002325A1 (en) * 2005-02-17 2007-01-04 Jimpei Tabata Fluorescence measurement apparatus
US7349093B2 (en) * 2005-02-17 2008-03-25 Matsushita Electric Industrial Co., Ltd. Fluorescence measurement apparatus
US20090080194A1 (en) * 2006-02-15 2009-03-26 Li-Cor, Inc. Fluorescence filtering system and method for molecular imaging
US20070188760A1 (en) * 2006-02-15 2007-08-16 Ahmed Bouzid Fluorescence filtering system and method for molecular imaging
US7286232B2 (en) * 2006-02-15 2007-10-23 Li-Cor, Inc. Fluorescence filtering system and method for molecular imaging
US11770503B2 (en) 2006-12-22 2023-09-26 Stryker European Operations Limited Imaging systems and methods for displaying fluorescence and visible images
US8498695B2 (en) * 2006-12-22 2013-07-30 Novadaq Technologies Inc. Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US20130286176A1 (en) * 2006-12-22 2013-10-31 Novadaq Technologies Inc. Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US9143746B2 (en) * 2006-12-22 2015-09-22 Novadaq Technologies, Inc. Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US11025867B2 (en) 2006-12-22 2021-06-01 Stryker European Operations Limited Imaging systems and methods for displaying fluorescence and visible images
US10694152B2 (en) 2006-12-22 2020-06-23 Novadaq Technologies ULC Imaging systems and methods for displaying fluorescence and visible images
US10694151B2 (en) 2006-12-22 2020-06-23 Novadaq Technologies ULC Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US20080239070A1 (en) * 2006-12-22 2008-10-02 Novadaq Technologies Inc. Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy
US20090095885A1 (en) * 2007-10-11 2009-04-16 Hager Harold E System and methods for detecting semiconductor-based photodiodes
US7709779B2 (en) * 2007-10-11 2010-05-04 The Boeing Company Method and apparatus for detecting an optical reflection indicative of a photodiode
US9642532B2 (en) 2008-03-18 2017-05-09 Novadaq Technologies Inc. Imaging system for combined full-color reflectance and near-infrared imaging
US10779734B2 (en) 2008-03-18 2020-09-22 Stryker European Operations Limited Imaging system for combine full-color reflectance and near-infrared imaging
US9814378B2 (en) 2011-03-08 2017-11-14 Novadaq Technologies Inc. Full spectrum LED illuminator having a mechanical enclosure and heatsink
US20150103229A1 (en) * 2013-02-13 2015-04-16 Panasonic Intellectual Property Management Co., Ltd. Multispectral imaging device and multispectral imaging method
US9843740B2 (en) * 2013-02-13 2017-12-12 Panasonic Intellectual Property Management Co., Ltd. Multispectral imaging device and multispectral imaging method
EP3100078A4 (en) * 2014-01-31 2017-09-06 JDS Uniphase Corporation An optical filter and spectrometer
US10190910B2 (en) 2014-01-31 2019-01-29 Viavi Solutions Inc. Optical filter and spectrometer
US9541750B2 (en) 2014-06-23 2017-01-10 Li-Cor, Inc. Telecentric, wide-field fluorescence scanning systems and methods
KR20160092085A (en) 2015-01-26 2016-08-04 이화여자대학교 산학협력단 Proactive portable algae detecting method and apparatus
KR20160131133A (en) 2015-05-06 2016-11-16 이화여자대학교 산학협력단 Control method for portable algae detecting apparatus and portable algae detecting apparatus
KR101683379B1 (en) 2015-05-29 2016-12-06 이화여자대학교 산학협력단 Portable algae detecting apparatus
US11237049B2 (en) 2015-08-05 2022-02-01 Viavi Solutions Inc. Optical filter and spectrometer
US10753793B2 (en) 2015-08-05 2020-08-25 Viavi Solutions Inc. Optical filter and spectrometer
US10048127B2 (en) 2015-08-05 2018-08-14 Viavi Solutions Inc. Optical filter and spectrometer
US10481100B2 (en) 2015-08-05 2019-11-19 Viavi Solutions Inc. In-situ spectral process monitoring
US9945790B2 (en) 2015-08-05 2018-04-17 Viavi Solutions Inc. In-situ spectral process monitoring
US11930278B2 (en) 2015-11-13 2024-03-12 Stryker Corporation Systems and methods for illumination and imaging of a target
US11298024B2 (en) 2016-01-26 2022-04-12 Stryker European Operations Limited Configurable platform
US10980420B2 (en) 2016-01-26 2021-04-20 Stryker European Operations Limited Configurable platform
CN107085291A (en) * 2016-02-15 2017-08-22 徕卡仪器(新加坡)有限公司 For the microscopical illumination filter system of multispectral fluorescence of performing the operation and its illumination wave filter
USD916294S1 (en) 2016-04-28 2021-04-13 Stryker European Operations Limited Illumination and imaging device
USD977480S1 (en) 2016-04-28 2023-02-07 Stryker European Operations Limited Device for illumination and imaging of a target
US10869645B2 (en) 2016-06-14 2020-12-22 Stryker European Operations Limited Methods and systems for adaptive imaging for low light signal enhancement in medical visualization
US11756674B2 (en) 2016-06-14 2023-09-12 Stryker European Operations Limited Methods and systems for adaptive imaging for low light signal enhancement in medical visualization
KR20170015959A (en) 2017-02-01 2017-02-10 이화여자대학교 산학협력단 Proactive portable algae detecting apparatus
US11140305B2 (en) 2017-02-10 2021-10-05 Stryker European Operations Limited Open-field handheld fluorescence imaging systems and methods
US10992848B2 (en) 2017-02-10 2021-04-27 Novadaq Technologies ULC Open-field handheld fluorescence imaging systems and methods

Also Published As

Publication number Publication date
US6441892B2 (en) 2002-08-27
EP1232387A4 (en) 2008-10-22
WO2001036948A1 (en) 2001-05-25
CN1409818A (en) 2003-04-09
CA2392228A1 (en) 2001-05-25
EP1232387A1 (en) 2002-08-21
JP2003515129A (en) 2003-04-22

Similar Documents

Publication Publication Date Title
US6441892B2 (en) Compact spectrofluorometer
US8130380B2 (en) Spectrometer and interferometric method
FI109149B (en) Spectrometer and method for measuring optical spectrum
US7738095B2 (en) Method and apparatus for compact spectrometer for detecting hazardous agents
US6091502A (en) Device and method for performing spectral measurements in flow cells with spatial resolution
US7084972B2 (en) Method and apparatus for compact dispersive imaging spectrometer
US8269174B2 (en) Method and apparatus for compact spectrometer for multipoint sampling of an object
US7440096B2 (en) Method and apparatus for compact spectrometer for fiber array spectral translator
US7548310B2 (en) Method and apparatus for compact spectrometer for multipoint sampling of an object
JP2511057B2 (en) Spectral analysis method and apparatus
US6323944B1 (en) Compact spectrofluorometer
JP6134719B2 (en) System and method for self-contrast detection and imaging of a sample array
US5615008A (en) Optical waveguide integrated spectrometer
US8130376B2 (en) Optical devices, spectroscopic systems and methods for detecting scattered light
US5422719A (en) Multi-wave-length spectrofluorometer
CA1229897A (en) Optics system for emission spectrometer
KR20130123389A (en) Systems and methods for detection and imaging of two-dimensional sample arrays
US10101273B2 (en) Optical emission collection and detection device and method
CN1089894C (en) High-resolution ratio, compacted intracavity laser spectrometer
US7352467B2 (en) Surface plasmon resonance imaging system and method
US20140085637A1 (en) Tunable light source system and method having wavelength reference capability
US20070127027A1 (en) Photometer having multiple light paths
CA2480463A1 (en) Hybrid-imaging spectrometer
JPH0219897B2 (en)
US20230236127A1 (en) Apparatus for determining the presence or concentration of target molecules

Legal Events

Date Code Title Description
AS Assignment

Owner name: JOBIN YVON, INC., NEW JERSEY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:XIAO, MING;REEL/FRAME:011818/0850

Effective date: 20010511

STCF Information on status: patent grant

Free format text: PATENTED CASE

AS Assignment

Owner name: HORIBA JOBIN YVON INC., NEW JERSEY

Free format text: CHANGE OF NAME;ASSIGNOR:JOBIN YVON INC.;REEL/FRAME:015312/0359

Effective date: 20040916

FPAY Fee payment

Year of fee payment: 4

FPAY Fee payment

Year of fee payment: 8

SULP Surcharge for late payment

Year of fee payment: 7

AS Assignment

Owner name: HORIBA INSTRUMENTS INCORPORATED, CALIFORNIA

Free format text: CHANGE OF NAME;ASSIGNOR:HORIBA JOBIN YVON INC.;REEL/FRAME:029173/0344

Effective date: 20111219

FPAY Fee payment

Year of fee payment: 12

SULP Surcharge for late payment

Year of fee payment: 11