|Publication number||US3661460 A|
|Publication date||9 May 1972|
|Filing date||28 Aug 1970|
|Priority date||28 Aug 1970|
|Also published as||CA936017A, CA936017A1, DE2142237A1, DE2142237B2, DE2142237C3|
|Publication number||US 3661460 A, US 3661460A, US-A-3661460, US3661460 A, US3661460A|
|Inventors||Elking Alan H, Groner Warren, Saunders Alex M|
|Original Assignee||Technicon Instr|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (10), Referenced by (100), Classifications (13), Legal Events (1)|
|External Links: USPTO, USPTO Assignment, Espacenet|
United States Patent [151 3,66 1,460 Elking et a1. May 9, 1972 54 METHOD AND APPARATUS FOR 3,504,183 3/1970 Salkowski et al. ..356/103 x OPTICAL ANALYSIS OF THE 3,515,884 6/1970 lmadate ..250/218 3,523,733 8/1970 Kling at al.... ...356/208 x CONTENTS OF A SHEATHED STREAM 3,560,754 2/1971 Kamentsky ..356/39 x Inventors: Alan H. Elking, White Plains; Warren Groner, Whitestone; Alex M. Saunders, Bedford Village. all of NY.
 Assignee: Technicon Instruments Corporation, Tarrytown, NY.
 Filed: Aug. 28, 1970 ] Appl. No.: 67,819
 U.S. Cl. ..356/36, 250/218, 250/222 PC, 356/102, 356/208, 356/246  Int.Cl. ..G01n1/00,G01n15/02,G01n2l/06  Field of Search ..356/36, 39-40, 356/102-104, 134, 201, 207-208, 246; 250/218, 222 PC [5 6] References Cited UNITED STATES PATENTS 2,731,877 1/1956 Clamann ..250/218 X 2,732,753 1/1956 OKonski ..356/103 X 2,875,666 3/1959 Parker et al.. ..250/218 X 2,920,525 1/1960 Appel et al... 356/102 X 3,398,286 8/1968 Ford et a1 ...356/103 X 3,440,866 4/1969 Ness et a1 ..356/39 X 14 PUMP T0 OVERFLOW 3) Primary Examiner-Ronald L. Wibert Assistant E.raminer-Warren A. Sklar Attorney-S. P. Tedesco and Rockwell S. E.
 ABSTRACT There is provided a method and apparatus for optical analysis by photometry of a substance flowing in a liquid stream within a coaxial sheath stream of a transparent liquid. The sheath stream, flowing in the same direction, entrains the inner stream so as to confine it concentrically. A photometer is used which includes a light source on one side of the sheathed stream in a position to direct light onto the inner stream which is cylindrical, the outer or sheath stream also being cylindrical. The photometer also includes a light detector externally of the sheathed stream in an angular position to detect the photometric results of impingement of light on the contents of the inner stream. Refraction of light at the interfaces of the sheath stream is compensated by varying the radius of the inner stream through the control of the flow of one stream with reference to the other.
The concept also includes the narrowing of the sheathed stream to a very small diameter in which it is confined by a wall structure in the area of examination.
19 Claims, 6 Drawing Figures 12 CONSTANT LEVEL RESERVOIR 40 SHEATH SOURCE 28 NEEDLE VALVE 0R RESISTOR LIGHT DETECTOR 26 VACUUM PUMP REGULATOR PATENTEUNNT 9 I972 3,661,460
SHEET 1 0T 3 FIG. 1
T4 PUMP To OVERFLOW\ P T 1 WWW LEVEL H RESERVOIR 28 i0 1 NEEDLE VALVE 0R SSOHUERACTEH RESISTOR LIGHT DETECTOR 2e I5 I f m ucNT l souRcE 25 QSw REGULATOR k E 2| LIQUID 22 TRAP INVENTORS ALAN H. ELKIND WARREN GRONER ALEX M. SAUNDERS ATTORNEY METHOD AND APPARATUS FOR OPTICAL ANALYSIS OF THE CONTENTS OF A SHEATI'IED STREAM BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to a method and apparatus for optical analysis by photometry of a substance flowing in a liquid stream within a coaxial sheath stream of a transparent confining liquid in which it is entrained.
2. Prior Art Fine streams utilized for optical examination of their content frequently clog in small diameter transparent tubing as is well known, especially in circumstances where the stream contains, intentionally or otherwise, particulate matter. Also, it has long been recognized that in the use of such fine tubing there is an undesirable pressure drop. I
At least certain of these difficulties were recognized by P. J. Crosland-Taylor who proposed, in an article entitled A Device for Counting Small Particles Suspended in a Fluid through a Tube", which appeared in the Jan. 3, 1953 issue of the publication NATURE, page 37, injecting a suspension of particles to be counted into a stream of liquid flowing in the same direction. Under conditions in which very little turbulence existed, the relatively wide column of particles was accelerated by the faster flowing stream to form a narrow column surrounded by the sheath liquid previously selected to match the index of refraction of the suspension.
The sheathed stream was enclosed in a block-like device for observation, and the author made no mention of any light refraction problem existing with reference to the outer interface of the sheath, probably, because in that device the viewing parts did not permit close observation of the sample. It presently appears that no thought was then given to the problem of refraction of light at both interfaces of a cylindrical sheath stream which encloses a cylindrical inner stream.
Subsequently, further experimental work was reported along the same general lines by P. F. Mullaney, M. A. Van Dilla, 1. R. Coulter, and P. N. Dean, in an article entitled "Cell Sizing: A Light-Scattering Photometer for Rapid Volume Determination which appeared in the publication THE REVIEW OF SCIENTIFIC INSTRUMENTS, Vol. 48, No. 8, Aug. 1969. A flow chamber was described in which the sheathed stream was injected into an ambient body of liquid confined by the chamber in the windowed observation area upstream of a restrictive orifice and carried therebeyond through the orifice. This work was based on the earlier work of P. J. Crosland-Taylor.
The later use by others of the last-mentioned flow chamber was fraught with optical problems among which was the preclusion of optics with close working distances, say, in the order of 2 mm, and multiple difficulties, same caused by entrapment of air bubbles, involving undesirable refraction of light in the observation area including but not limited to refraction of light at the interfaces of the sheath stream. Also, observation was restricted to a small area, upstream of the aforementioned restrictive orifice.
The present invention effectively tends to obviate the aforementioned difficulties, particularly in dark field light-scattering photometric techniques.
SUMMARY OF THE INVENTION It is an object of the invention to provide an improved method and apparatus for optical analysis by photometry of a substance flowing in a liquid stream within a coaxial sheath stream of a transparent liquid which sheath stream flows in the same direction and entrains the inner stream so as to confine it in concentric relationship.
Another object includes the narrowing of the sheathed stream to a very small diameter in which it may be confined by a wall structure in the area of examination, and the avoidance in this area of any more volume of the sheathed stream than necessary to carry the sample. This tends effectively to reduce any ambient fluid which might obscure the sample.
A further object is to provide equal and opposite refraction of light in photometric analysis of a cylindrical sheathed stream by varying the radius of the inner stream within limited but practical ranges through the control of the relative flow of the inner and outer streams.
According to the invention, there may be provided in such apparatus a flow chamber having an elongated passageway portion provided with an outlet end and at the other end a tube inlet of substantially smaller diameter extending concentrically a distance into the passageway portion and terminating forwardly in a free tube end. The'flow chamber further comprises a second inlet in the passageway portion at a location a distance rearwardly from the free tube end. The flow cell further comprises a restriction in the aforementioned passageway portion intermediate the aforementioned outlet end thereof and the free tube end, which defines a round opening, the internal surfaces of the passageway portion of the tube being circular.
Provision is made for flowing a sample stream through the tube inlet into the passageway portion and for flowing a sheath stream into the aforementioned second passageway inlet at a greater velocity than the sample to entrain the latter so as to increase the velocity of the sample stream and narrow it as the streams flow toward the aforementioned restriction of the passageway portion wherein they are narrowed proportionately, thereby both being accelerated.
A photometer is provided downstream from the upstream extremity of the restriction of the passageway portion and includes a light source, on one side of the sheathed stream in a position to direct light'onto the sample stream, and a light detector externally of the sheathed stream in an angular position to detect the photometric results of impingement of light on the contents of the sample stream. A further provision is made for controlling the relative flow of the streams which controls the radius of the sample stream, so that the last named radius may be varied and thereby compensate for refraction of light at the interfaces of the sheath stream.
BRIEF DESCRIPTION OF THE DRAWING In the drawing:
FIG. 1 is a diagramatic view of apparatus for optical analysis by photometry including a flow chamber, embodying the invention;
FIG. 2A is a view of the flow chamber of FIG. 1 illustrating it in longitudinal horizontal section;
FIG. 2B is a longitudinal elevational sectional view of the flow chamber;
FIG. 3 is an enlarged sectional view taken on line 3-3 of FIG. 2B, illustrating the sheathed stream within the flow chamber;
FIG. 4 is a further enlarged, fragmentary view in section taken on line 4--4 of FIG. 2B, illustrating the sheathed stream within the flow chamber and indicating how refraction of light at the interfaces of the sheath stream is compensated; and
FIG. 5 is a view similar to FIG. 2A illustrating in vertical section a modified form of the flow chamber.
DESCRIPTION OF THE PREFERRED EMBODIMENT In FIG. 1 there is shown the principal elements of apparatus for optical analysis by photometry of a substance flowing in a liquid stream within a coaxial sheath stream of a transparent liquid, the illustration being by way of example only. There is indicated at 10 a supply of transparent liquid for the sheath stream, the source being illustrated as a flask from which passes tube 11 emptying into an elevated vessel 12 for the sheath stream. The vessel 12 has an overflow outlet including a tube 13 to carry off excess liquid and return it to the source 10 by gravity.
A pump 14 is provided to convey liquid from the source 10 to the vessel 12 which forms a constant-level reservoir for the sheath stream liquid. The bottom of the vessel is provided with a liquid outlet connected to a gravity feed tube 15 connecting the vessel 12 with a lower flow chamber indicated generally at 16. The flowcharnber is gen'erally'of elongated tubular shape and, in the illustrated form, the tube 15 is branched so that the flowchamber hasa pair of liquid inlets 17 for the sheath stream which are arranged opposite one another in the side-.
rninating forwardly .a distance within the longitudinal passageway of the chamber 16 in a free end. The tube 18 is ar- 3, 1 960 without causing undue pulsationsofthe sample within ranged concentrically 'with reference to the longitudinal passageway of theflow chamber, the tube 18 being of substantially smaller outer diameter than the which it extends. v v v The other'end of the flow chamber 16 is a discharge end passageway portion into connected to a tube 19 discharging into a suitable receptacle which-is here shown as a stoppered flask 20. Vacuum tube 21 is connect'ed. to the upper portion of the flask 20 to create a vacuumfor the purpose of withdrawing the sheathedstream from the flow chamber 16, andto this end the tube 21 is con- ;nected to. a vacuum pump 22 through a regulator 23, and the tube 21' isprovided with a suitable pressure guage24. v
The flow chamber 16, details of which will be discussed hereinafter with reference to other views of the drawings, is structured largely of a transparent material such as a suitable glass, and has associated with a transparent region thereof a photometer including a light source 25 at one sideof the flow chamber and at the opposite'sidea'light detector 26 so that light from the sourcepasses through the-contents'of the flow chamber to the light detector in the illustrated form.
This arrangement, however,'is not a limitation. In this connection it may be noted that while the light source and light detector are indicated to be in opposing relationship, the light detector might be arranged at right angles to the path of light impinged on the sheathed stream todetect fluoresence, for example, in the contents of the flow chamber 16. In a certain photometric technique, the light emitted from the light source andimpinged on the stream to be analyzed may be reflected backin the direction of the light source for a distance before being diverted vto the light detector convenientlylocated to receive such diverted light rays, and such a technique is also possible according to the invention. The illustrated arrange 'ment of the light source and light detector permits analysis of a sample by its light transmitting characteristics or its light absorbing characteristics,
techniques. Y I v For illustrative purposes it may be considered that the sample contains particles in suspension to be counted by a forward scattering of lightby the particles, that is, toward the light detector in the position shown, as they pass substantially one Thesample stream may be fed to the inlet tube 18 intermittently or continuously and be pulled or pushed for introduc-- tion into the flow chamber. U.S. Pat. No. 2,879,14l issued and; also by. light-scattering desired the sheath; stream, like thesample stream, may be pushed or, pulled by a pump for introduction into the flow inlet forthe sheath streamf, V. v 7
:It will be apparent that theflow chambermay haveasingle inlet for the sheath stream, such as one concentric with .the tube 18. lt is. necessary, here, that the sheath streamhave a sufficient distance of travel fromjts point of entry to the free forward end of 'the sample tube l 8-'to completely fill the passageway portion and establishsteady state laminar flow surrounding the tube in order to properly entrain the sample stream injectedthereinto from .the'tube 18 on its passage toward the discharge end of the flow-chamber connected to chamber inlets 17, which may be considered together as an the tube 19, as will more readily apparent fromthe discussion hereinafter of other views of the drawings. 1
As will appear more fully hereinafter, the entrained sample stream is reduced in diameter after leaving the tube 18, and the radius of the sample stream may be variedby-controlling the relative flow of the sample and sheath streams as previ ously indicated. As it is usually deemed undesirable to place a restrictive device'such as a valve or the like in the sample stream inlet line due to the riskof clogging. of thesample stream, a needle valve 28 maybe interposedin the sheath stream tube 15 intermediate the vessell2 and the chamber inlet 17 tocontrol the inlet flowof the sheath stream'to the flow chamben-Aresistor may be used in place of the valve 28, ifv desired. It will .be evident thatif the flow of the sheath stream is reduced, theradius of the sample stream is enlarged. Also, if the sheath-stream flow is' increased, the radius of the sample stream. diminishes. Refraction of light impinged on the sample stream by thelight source 25 is a" function, atleastin part, of the radius of the sample stream. v
Referring now to'the details of theflow chamber 16 shown in FIGS. 2A though 4, the chamber includes an elongated tubular body 30 of transparent glass material having diametrically opposite .side' arms in alignment with one another providing the inlets 17 for the sheath'stream, in which arms nipple fittings may be received, respectively, for connection to the flow chamber and are of approximately the: same internal after another in a stream intermediate the light source and the Mar. 24, 1959 discloses-(the disclosure'of which is incor-U porated herein by reference) a suitable automated sample supply apparatus, not shown, which has provision for holding a plurality of samples, which may be used. The samples of a se- I ties in tubing may be separated from one another by a seg:
menting fluid which maintains the integrity of the samples and tends to cleanse the tubing wall as the sample stream passes toward the examination area as described in the last-mentioned patent. Usually any segmenting fluid is removed prior to entry of a smple into the flow chamber as by a device, not
shown, such as that illustrated and described in US. Pat.,No.
3,109,714 issued Nov. 5, 1963.1t has been found that samples.
may be conveniently transmitted to and through the last.-' named device by a peristaltic pump, not shown, such as that illustrated and described in 0.8. Pat. No. 2,935,028 issued May diameter as the longitudinal portion 32 of the flow chamber passageway with which they communicate,-which maybe approximately .125 inch.
. The longitudinal passageway portion 32 continues tothe rear end of the flow chamber and is closed by a plug 34 through the center of which, extends'the mid-portion of the sample stream inlet tube 18, so that the tube Isis concentrically supported in fluid-tight relation thereby in thepassageway portion 32. The forward free end 36 of the tube 18 extends beyond the inlet arms 17 and terminates in the passagewayportion 32 whichis circular in cross section. The cylindricaltube 18 which has an outer diameter substantially smaller than that Of the passageway portion 32 may have an inner diameter of approximately 0.0195 inch and be con- 7 stituted by hypodermic tubing or of a small-diameter tube structure of a non-corrosive metal. lts'fluid passageway is circular in cross section.
As shown in nos. 24 and 2B, therear end portion of thetube 18 is received in one end-of a sleeve '37 the other end of which sleeve mayreceive a nipple fitting 38 for connection to v a suitable sample tube not shown. A distance forwardly of the free end 36 of the tube 18, the inner wall surface of the longitudinal passageway in the tubular body 30 slopingly narrows creating a restriction, indicated at 42, of circular cross section defining an orifice and extending forwardly in the area of and beyond the light path between the light source 25 and the light detector 26 as shown in FIGS. 2A and 2B.
In the region of the light path, outer wall surfaces of the tubular body are thinned and flattened, as at 44, on opposite sides thereof in opposing relation to the light source and the light detector to minimize light refraction at the outer surface of the tubular body 30 and to permit the aforementioned optical elements 25, 26 to closely approach the fluid contents of the restricted passageway portion 42, as indicated in FIGS. 2A and 3. This close working distance of the optical elements from the fluids contents may be approximately 0.010 inch, and permits use of a high numerical aperature condenser and objective. If desired, the portion of the tubular body 30 of the flow chamber in the light path may be immersed in optical oil for a refractive index match.
The restriction 42 in the tubular body 30, which may be 0.010 inch, is illustrated as extending through the thickened discharge end 40 (FIG. 2A) of the body 30 received in one end of a sleeve 41 the other end of which receives a nipple fitting 43 for connection to the discharge tube 19 (FIG. 1) for discharge of the composite liquid stream from the flow chamber into the waste receptacle or trap 20 shown in FIG. 1. The illustrated vacuum pump 22 connected through tube 21 to the receptacle 20 effectively discharges the flow chamber, and it has been found that the pump 22 may be run at varying vacuums without changing the radial dimension of the sample stream within the sheath stream in the examination area provided by the restricted passageway portion 42 of the flow chamber. 4
It will be understood from the foregoing that when the sample stream is injected through the tube 18 into the passageway portion 32 of the flow chamber it is entrained and accelerated by the faster flowing laminar sheath stream, and the streams are proportionately narrowed and accelerated as they are influenced by the restriction 42 of the passageway prior to reaching the examination area. Hence, small particles in diluted suspension forming the sample stream tend to follow one another through the passageway portion 42 rather than pass abreast of one another, which enables the particles to be counted by the action of the photometer which operates a suitable recorder not shown. In FIGS. 3 and 4 the sample stream is indicated at 46 and the sheath stream at 48.
The diameter of the sample stream may be, in a particular instance, depending on the relative flow of the sample and sheath streams, approximately 0.003 inch. Also, by way of example, the sample stream may equal 9 per cent of the total flow. In such circumstances a total flow of the combined streams through the flow chamber of 5 ml per minute, or 3 millionths of a cubic foot per second, equal a Reynolds number of5 10.
In the modified form of the invention illustrated in FIG. 5, the flow chamber, indicated generally at 50, has an elongated block-like body 51 vertically arranged provided through its upper end with a pair of converging bores or sheath stream inlets 52 respectively receiving nipple fittings 54 for connection to the respective branches of a sheath stream supply tube similar to the tube previously described with reference to FIG. 1. These inlets 52 converge, as shown, in a central, longitudinal bore or passageway portion 56 of the body 51, extending through the lower end thereof. Through the upper end portion of the body 51, there is provided a central longitudinal bore intermediate the sheath stream inlets 52 which bore, communicating with the passageway portion 56, receives a sample stream inlet tube 58, similar to the sample stream inlet tube 18, previously described. The tube 58 extends through a supporting plug 60, provided in an enlargement of the lastmentioned bore, and the tube 58 is concentrically arranged in the passageway portion 56, with its lower free end 62 terminating a distance upwardly from the lower end of the passageway portion 56, as illustrated in F IG. 5.
As shown in the last-mentioned view, a restriction is formed in the lower discharge end of the passageway portion 56, the restriction in this instance being formed by a plug 64 inserted for support in the lower end of the passageway portion 56, the plug 64 having a central opening 66 defined by a funnel-like surface. From the opening 66 in the plug 64, the sheathed stream is injected downwardly into the ambient atmosphere for examination by a photometer, illustrated as comprising a light source 68, closely located at one side of the stream, and the light detector 70, located closely to the stream on the diametrically opposite side thereof, for a light path therebetween. Also, as shown in FIG. 5, there may be provided a funnel 72 to catch the streams shortly after they cross the last mentioned light path and collect them for drainage into a suitable receptacle therebelow, not shown.
In the operation of the flow chamber 50, the sheath liquid supplied to the inlets 52 of the flow chamber 50 picks up, in the passageway portion 56, the sample stream injected into the passageway portion 56 through the tube 58, in a manner to entrain it and accelerate the sample stream, narrowing it as the streams flow toward the restriction formed in the passageway portion 56 by the plug 64, wherein they are narrowed proportionately, thereby both being accelerated, all in a manner similar to the operation of the flow chamber 16 previously described. As previously indicated, the sheathed stream is unconfined after it exits from the flow chamber 50 through the plug 64 and passes across the light path between the light source 68 and the light detector 70. This makes possible extremely short working distances from the sheathed stream to the optical elements 68, 70, with attendant apparent advantages.
To compensate for the refraction of light at the interfaces of the sheath stream in the area in which the stream is examined, the radius of the inner or sample stream may be varied, as in the use of the previously described flow chamber 16, by adjusting the relative flow of the sheath and sample streams. This control of the relative flow of the streams may be accomplished in the previously described manner. Another advantage of the form of the invention shown in FIG. 5 over that shown in FIG. 1, is that by injecting the sheathed stream into the ambient atmosphere for examination, a significant pressure drop may be avoided in the area of examination.
However, one of the advantages of the form of the invention shown in FIG. 1, is that the flow chamber 16 may be oriented in any position so that the optical elements associated with the flow chamber may accordingly be positioned, that is, vertically or horizontally. Ifthe flow chamber 16 is arranged vertically with the flow directed upward, it is obvious that any air bubbles which might be caught in the chamber will be swept to the upper end thereof and out of the viewing area. Still another advantage of the form of FIG. 1, is that the sheathed stream is confined in the examination area so as to prevent splattering of the optical elements associated with the flow chamber as by an aberration in the sheathed stream.
Referring now to the optical problems, specifically refraction of light, overcome by the invention, it is known that the external surface of a cylinder such as the sheath stream will refract light if the substance with which it is in contact has a different index of refraction, such as air or glass. This refraction may be compensated within workable limits including the indexes of refraction of the outer substance such as air or glass, the sheath stream and the sample stream.
Compensation is achieved if rays 74 (FIG. 4) parallel to the optical axis 75 reach the center line of the sample stream in parallel relation to the axis 75 of light from the aforementioned light source 25. This compensation may be obtained by determining and establishing the correct radius of the sample stream with reference to the particular operating conditions prevailing at the time including the last-mentioned indexes of refraction. The radial dimension of the sample stream must be such as to bring about equal and opposite refraction at the two interfaces of the sheath stream.
In the form illustrated in FIG. 4 in which the sheathed stream is enclosed within a hollow cylindrical wall surface, the index of refraction N, of the sample stream must be greater than the index of refraction N, of the sheath stream to obtain compensation of refraction at the interfaces of the sheath stream. The converse is true in the situation of FIG. 5 in which the sheath stream in the area of examination is unconfined,
streamsis less than the absolute difference in refraction indexes of the outer stream and that (here, glass) to which it is externally exposed.
Thus,,in the following an example, where N is 1.51; N, is l .36; N is approximately 1.38; and r,, the radius of the sheath stream, is 0.005 inch: in solving for r, the radius of the sample stream, the following equation is used:
1 r=a p proximately .00076 inch from elementary optics wherein:
' In the foregoing example, the sample stream is constituted by human. blood diluted with propylene glycol and in which the red cells are hemolyzed. The red cell ghosts have the same index of refraction as the diluent so that the-red cells are rendered invisible, for the counting by photometric means, as aforesaid, of the white cells. One obvious advantage of the invention is that there is no requirement in the use of the flow chamber that the index of refraction of the sheath stream match the index of refraction of the sample stream.
The photometer has a conventional ocular, not shown, to observe, with the human eye, the flow of the stream in the flow chamber so that it may be determined whether or not refractive errors have been compensated. in this connection a conventional target, not shown, of the optical elements, projected through the flow chamber is brought into sharp focus at the particular sample stream radius which achieves compensation of light refraction as indicated above. The photometric viewing area should be larger than the diameter of the sample team.
It is believed the many advantages of this invention will now be apparent to those skilled in the art. Theforegoing description is illustrative, rather than limiting, as a number of variations and modifications may be made without departing from the true spirit and scope of the invention. The invention is limited only to the scope of the following claims.
What is claimed is:
1. Apparatus for optical analysis of a sample stream including a liquid in a coaxial sheath stream liquid comprising: means defining a flow chamber. having an elongated passageway portion having a forward outlet end and at a rear end of said portion a tube inlet of substantially smaller diame ter than said passageway portion extending concentrically a distance forwardly into said passageway portion and terminating forwardly in a free tube end, the last-named means defining a second inlet in said-passageway portion at a location a distance rearwardly from said free tube end, and the lastnamed means also defining a restriction with a narrow sloping approach in said passageway portion intermediate said outlet end thereof and said free tube end, the internal andexternal surfaces of said free tube end and the internal surface of said passageway portion being circular in cross section, means for flowing a sample stream through said'tube inlet into said passageway portion, means for flowing a sheath stream into said second passageway inlet at a greater flow rate than the sample stream to entrain the latter so" as to increase the velocity of the sample stream and narrow it as the streams flow toward said restriction of the passageway portion, photometric means downstream from the upstream extremity of restriction of the passageway portion in proximity to the sheathed stream and including a light source on one side of the sheathed stream in a position to direct lightonto the contents of the sample stream anda light detector externally of the sheathed stream in angular position to detect the photometric results of impingement of light on the sample stream, and means controlling therelative flow rates of said streams into said flow chamber, which controls the radius of the sample I stream, so that in the area where said light is directed on the sample stream the last-named radius may be varied proportionately to the radius of the inner surface of the sheath stream for equal and opposite refraction of light at the interfaces of said sheath stream. v
2. Apparatus as defined in claim 1, wherein: said means controlling the relative flow rates of said streams comprises a device operative to restrict the flow of the sheath stream into the flow chamber.
3. Apparatus as defined in claim 1, wherein: said means for flowing the sheath stream into said flow chamber comprises a constant-level reservoir establishing a head of sheath stream liquid.
4. Apparatus as defined in claim 1, wherein: said means for flowing the sample stream into said tube inlet of the flow chamber comprises a pump upstream of the tube inlet.
5. Apparatus as defined in claim 1, wherein: said outlet end of the flow chamber is connected to a pump.
GsApparatus asdefined in claim 1, wherein: said means controlling the relative flow rates of the streams comprises a valve operative to vary the flow of the sheath stream into the flow chamber. v
7 Apparatus as defined in claim 1, wherein: said means for flowing the sample stream into said sample tube inlet of the flow chamber comprises a pump upstream of the tube inlet, said outlet end of the flow chamberbeing connected to a liquid trap throughan outlet conduit, and further comprising a vacuum pump 'operatively connected to the last-named outlet conduit.
V 8. Apparatus as defined in claim 1, wherein: the flow chamber is vertically arranged.
9. Apparatus as-defined in claim 1, wherein: the sheathed stream is directed from said outlet end of the flow chamber into a gas, and said photometric means examines the sheathed sample stream in said gas.
10. Apparatus as defined in claim Lwherein: the sheathed stream is directed into ambient air from said outlet end of the flow chamber, and said photometric means examines the sheathed sample stream in said ambient air.
11. Apparatus as defined inv claim 1, wherein: the flow chamber is horizontally arranged.
12. Apparatus as defined in claim 1, wherein: said means defining a flow chamber comprises a tubular body having said restriction of the passageway portion therein.
13. Apparatus as defined in claim .1, whereimsaid means defining a flow chamber comprises a tubular body having said restriction of the passageway portion therein, said restriction enclosing the sheathed stream where light from said source. is impinged on the sample stream.
14. A method for optical analysis of a sample stream including a liquid in a coaxial sheath stream liquid comprising: providing means defining an elongated passageway portion of circular cross section in a flow chamber which portion has at one end thereof a circular restriction; flowing a first laminar liquid stream into said passageway portion in a direction toward said restriction; concurrently flowing in the same direction a slower moving liquid stream of cylindrical cross section and of a substantially smaller diameter than said first stream, containing a sample, into the first stream concentrically so that the latter sheaths and entrains the second stream, accelerating and narrowing it as the streams approach said restriction in a concentric relation to one another; directing an impinging light beam of a photometer, in a direction transversely of the aforementioned direction of flow, onto the samplecontaining stream for photometric analysis of the sample by a light detector of the photometer, downstream from the upstream extremity of said restriction; and controlling the relative flow rates of the streams into said passageway portion, which controls the radius of the second or sample stream, so that in the area where said light beam impinges said sample stream the last-named radius may be varied proportionately to the radius of the inner surface of the sheath stream for equal and opposite refraction of light at the interfaces of the sheath stream.
15. The method as defined in claim 14, wherein: the step of controlling the relative flow of the streams into said flow chamber comprises controlling said sheath stream.
16. The method as defined in claim 14, wherein: the step of flowing the sheath stream into the flow chamber comprises flowing the sheath stream from a constant-pressure source.
17. The method as defined in claim 14, further comprising introducing the sheath and sample streams into the How chamber under positive pressure, and withstanding the sheath stream from the flow chamber under negative pressure through an outlet of the flow chamber downstream from said restriction.
18. The method as defined in claim 14, further comprising narrowly confining the sheath stream where said light beam is impinged on the sample stream.
19. The method as defined in claim 14, further comprising directing the sheathed stream into a gas from the flow chamber in a downward direction and examining the sheathed stream by the photometer in said gas.
" i t 8 t I UNITED STATES PATENT OFFICE CERTIFICATE :OF CORREC' IICPN Patent No. 3 1,460 Dated May 9, 1972 Inventor(s) Alan H- -kind Qt a1 It is certified that error appears in the above-identified patent andlthat said Letters Patent I are hereby corrected as shown below:
0n the cover Sheet  the inventor's name "Alan H. 'E lking" should read Alan H. Elkind Signed and sealed this 7th day of Novembervl972';
EDWARD M.FLEI'CHER,JR. ROBERT GOTTSCHAIK Attesting Officer Commissioner of Patents )RM p' v USCOMM-DC 6O376-P69 ll 5. GOVERNMENT PRINTING OFFICE; I96? 0-386-33
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US2731877 *||20 Jul 1953||24 Jan 1956||clamann|
|US2732753 *||16 Apr 1948||31 Jan 1956||o konski|
|US2875666 *||13 Jul 1953||3 Mar 1959||Ohio Commw Eng Co||Method of simultaneously counting red and white blood cells|
|US2920525 *||13 Aug 1956||12 Jan 1960||Arthur V Appel||Apparatus for automatically counting and sizing airborne particles|
|US3398286 *||17 Jul 1964||20 Aug 1968||Union Carbide Corp||Radiation sensitive evaporative analyzer|
|US3440866 *||1 Sep 1966||29 Apr 1969||Research Corp||Prothrombin timer apparatus and method|
|US3504183 *||12 Sep 1966||31 Mar 1970||Iit Res Inst||Particle counter using a variable test volume|
|US3515884 *||26 Apr 1968||2 Jun 1970||Toa Electric Co Ltd||Detecting and counting apparatus for particles suspended in a liquid|
|US3523733 *||5 Jan 1966||11 Aug 1970||Technicon Corp||Method and apparatus for particle counting|
|US3560754 *||17 Nov 1965||2 Feb 1971||Ibm||Photoelectric particle separator using time delay|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US3861800 *||27 Jul 1972||21 Jan 1975||Becton Dickinson Co||Particle counter independent of flow rate|
|US3871770 *||4 Jun 1973||18 Mar 1975||Nuclear Data Inc||Hydrodynamic focusing method and apparatus|
|US3893766 *||14 Jun 1973||8 Jul 1975||Coulter Electronics||Apparatus for orienting generally flat particles for slit-scan photometry|
|US4110043 *||24 Sep 1976||29 Aug 1978||Gesellschaft Fur Strahlen- Und Umweltforschung Mbh, Munchen||Apparatus for counting and classifying particles|
|US4284412 *||13 Jul 1979||18 Aug 1981||Ortho Diagnostics, Inc.||Method and apparatus for automated identification and enumeration of specified blood cell subclasses|
|US4408877 *||10 Apr 1980||11 Oct 1983||Ernst Leitz Wetzlar Gmbh||Device for hydrodynamic focussing of a particle-suspension in a liquid flow cytophotometer|
|US4565448 *||11 Mar 1983||21 Jan 1986||E. I. Du Pont De Nemours And Company||Particle counting apparatus|
|US4606631 *||4 May 1984||19 Aug 1986||Kabushiki Kaisha Toshiba||Particle counter|
|US4781459 *||22 Apr 1986||1 Nov 1988||Horiba, Ltd.||Apparatus for measuring the amount of minute particles contained in liquid|
|US4971912 *||14 Jul 1987||20 Nov 1990||Technicon Instruments Corporation||Apparatus and method for the separation of immiscible liquids|
|US4983038 *||7 Apr 1988||8 Jan 1991||Hitachi, Ltd.||Sheath flow type flow-cell device|
|US5046853 *||12 Jun 1989||10 Sep 1991||Basf Aktiengesellschaft||Measuring the degree of dispersion in flowing suspensions|
|US5194909 *||4 Dec 1990||16 Mar 1993||Tycko Daniel H||Apparatus and method for measuring volume and hemoglobin concentration of red blood cells|
|US5370842 *||20 Nov 1992||6 Dec 1994||Canon Kabushiki Kaisha||Sample measuring device and sample measuring system|
|US5593564 *||24 Jul 1995||14 Jan 1997||Hewlett-Packard Company||Microcolumn-microcolumn flow interface and method|
|US5602039 *||14 Oct 1994||11 Feb 1997||The University Of Washington||Flow cytometer jet monitor system|
|US5602349 *||14 Oct 1994||11 Feb 1997||The University Of Washington||Sample introduction system for a flow cytometer|
|US5643796 *||14 Oct 1994||1 Jul 1997||University Of Washington||System for sensing droplet formation time delay in a flow cytometer|
|US5726364 *||10 Feb 1997||10 Mar 1998||The University Of Washington||Sample introduction apparatus for a flow cytometer|
|US5819948 *||21 Aug 1997||13 Oct 1998||Van Den Engh; Gerrit J.||Particle separating apparatus and method|
|US5882599 *||15 Mar 1995||16 Mar 1999||Counting Technology Limited||Diluter|
|US6133044 *||16 Apr 1996||17 Oct 2000||University Of Washington||High speed flow cytometer droplet formation system and method|
|US6248590||27 Feb 1998||19 Jun 2001||Cytomation, Inc.||Method and apparatus for flow cytometry|
|US6507400||19 Feb 2000||14 Jan 2003||Mwi, Inc.||Optical system for multi-part differential particle discrimination and an apparatus using the same|
|US6589792||26 Feb 1999||8 Jul 2003||Dakocytomation Denmark A/S||Method and apparatus for flow cytometry|
|US6646742||19 Feb 2000||11 Nov 2003||Mwi, Inc.||Optical device and method for multi-angle laser light scatter|
|US6750060||25 Mar 2003||15 Jun 2004||Sysmex Corporation||Sheath liquid for particle analyzer|
|US6819411||2 Feb 1998||16 Nov 2004||Xy, Inc.||Optical apparatus|
|US6861265||12 Oct 2000||1 Mar 2005||University Of Washington||Flow cytometer droplet formation system|
|US7012689||17 May 2002||14 Mar 2006||Dako Colorado, Inc.||Flow cytometer with active automated optical alignment system|
|US7024316||20 Oct 2000||4 Apr 2006||Dakocytomation Colorado, Inc.||Transiently dynamic flow cytometer analysis system|
|US7094527||29 Nov 2001||22 Aug 2006||Xy, Inc.||System for in-vitro fertilization with spermatozoa separated into X-chromosome and Y-chromosome bearing populations|
|US7169548||9 Jan 2003||30 Jan 2007||Xy, Inc.||Sperm cell processing and preservation systems|
|US7195920||25 Feb 2003||27 Mar 2007||Xy, Inc.||Collection systems for cytometer sorting of sperm|
|US7208265||5 Jan 2000||24 Apr 2007||Xy, Inc.||Method of cryopreserving selected sperm cells|
|US7221453||16 Nov 2004||22 May 2007||Xy, Inc.||Optical apparatus|
|US7242474 *||27 Jul 2004||10 Jul 2007||Cox James A||Cytometer having fluid core stream position control|
|US7256882 *||21 Jun 2004||14 Aug 2007||Bgi Instruments, Inc.||Photometer device and method|
|US7371517||9 May 2001||13 May 2008||Xy, Inc.||High purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa|
|US7586604||22 May 2007||8 Sep 2009||Xy, Inc.||Optical apparatus|
|US7618770||17 Nov 2009||Xy, Inc.||Methods and apparatus for reducing protein content in sperm cell extenders|
|US7629113||8 Dec 2009||Xy, Inc||Multiple sexed embryo production system for bovine mammals|
|US7713687||29 Nov 2001||11 May 2010||Xy, Inc.||System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations|
|US7723116||25 May 2006||25 May 2010||Xy, Inc.||Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm|
|US7758811||20 Jul 2010||Inguran, Llc||System for analyzing particles using multiple flow cytometry units|
|US7760351||20 Jul 2010||Honeywell International Inc.||Cytometer having fluid core stream position control|
|US7771921||28 Sep 2006||10 Aug 2010||Xy, Llc||Separation systems of frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations|
|US7772005||29 Jul 1999||10 Aug 2010||Xy, Llc||Method of establishing an equine artificial insemination sample|
|US7799569||16 Mar 2009||21 Sep 2010||Inguran, Llc||Process for evaluating staining conditions of cells for sorting|
|US7820425||26 Oct 2010||Xy, Llc||Method of cryopreserving selected sperm cells|
|US7833147||22 Jul 2005||16 Nov 2010||Inguran, LLC.||Process for enriching a population of sperm cells|
|US7838210||23 Nov 2010||Inguran, LLC.||Sperm suspensions for sorting into X or Y chromosome-bearing enriched populations|
|US7855078||15 Aug 2003||21 Dec 2010||Xy, Llc||High resolution flow cytometer|
|US7892725||22 Feb 2011||Inguran, Llc||Process for storing a sperm dispersion|
|US7923252||12 Apr 2011||University Of Washington||Droplet formation systems for flow cytometers|
|US7929137||8 Sep 2009||19 Apr 2011||Xy, Llc||Optical apparatus|
|US7943384||7 Jun 2010||17 May 2011||Inguran Llc||Apparatus and methods for sorting particles|
|US8137967||21 Aug 2006||20 Mar 2012||Xy, Llc||In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations|
|US8211629||1 Aug 2003||3 Jul 2012||Xy, Llc||Low pressure sperm cell separation system|
|US8486618||18 Jul 2011||16 Jul 2013||Xy, Llc||Heterogeneous inseminate system|
|US8497063||24 Aug 2010||30 Jul 2013||Xy, Llc||Sex selected equine embryo production system|
|US8652769||9 Aug 2010||18 Feb 2014||Xy, Llc||Methods for separating frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations|
|US8664006||1 Mar 2013||4 Mar 2014||Inguran, Llc||Flow cytometer apparatus and method|
|US8709817||7 Feb 2013||29 Apr 2014||Inguran, Llc||Systems and methods for sorting particles|
|US8709825||1 Mar 2013||29 Apr 2014||Inguran, Llc||Flow cytometer method and apparatus|
|US8748183||7 Feb 2013||10 Jun 2014||Inguran, Llc||Method and apparatus for calibrating a flow cytometer|
|US8869594 *||4 Oct 2012||28 Oct 2014||Azbil Corporation||Particle detecting device evaluating system and particle detecting device evaluating method|
|US9040304||12 Mar 2014||26 May 2015||Inguran, Llc||Multi-channel system and methods for sorting particles|
|US9134220||27 Jul 2005||15 Sep 2015||Beckman Coulter, Inc.||Enhancing flow cytometry discrimination with geometric transformation|
|US9145590||1 May 2008||29 Sep 2015||Xy, Llc||Methods and apparatus for high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa|
|US9365822||11 Feb 2013||14 Jun 2016||Xy, Llc||System and method for sorting cells|
|US9377390||10 Apr 2015||28 Jun 2016||Inguran, Llc||Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm|
|US20020096123 *||12 Jun 2001||25 Jul 2002||Colorado State University, Colorado State University Research Foundation||Integrated herd management system utilizing isolated populations of X-chromosome bearing and Y-chromosome bearing spermatozoa|
|US20030129091 *||25 Feb 2003||10 Jul 2003||Colorado State University Through Its Agent, Colorado State University Research Foundation||Collection systems for cytometer sorting of sperm|
|US20030211009 *||18 May 2001||13 Nov 2003||Buchanan Kris S.||Rapid multi-material sample input system|
|US20040053243 *||9 May 2001||18 Mar 2004||Evans Kenneth M.||High purity x-chromosome bearing and y-chromosome bearing populations of spermatozoa|
|US20040055030 *||9 Jan 2003||18 Mar 2004||Xy, Inc.||Sperm cell processing and preservation systems|
|US20050110996 *||16 Nov 2004||26 May 2005||Sharpe Jonathan C.||Optical apparatus|
|US20050153458 *||28 Feb 2005||14 Jul 2005||University Of Washington||Droplet formation systems for flow cytometers|
|US20050280820 *||21 Jun 2004||22 Dec 2005||Gussman Robert A||Photometer device and method|
|US20060023207 *||27 Jul 2004||2 Feb 2006||Cox James A||Cytometer having fluid core stream position control|
|US20060259253 *||31 Mar 2006||16 Nov 2006||Dako Colorado, Inc.||Systems for transiently dynamic flow cytometer analysis|
|US20070026378 *||2 Sep 2005||1 Feb 2007||Xy, Inc.||Methods and apparatus for reducing protein content in sperm cell extenders|
|US20070099171 *||7 Dec 2006||3 May 2007||Xy, Inc.||Sperm Suspensions for Sorting Into X or Y Chromosome-bearing Enriched Populations|
|US20070099260 *||20 Dec 2006||3 May 2007||Xy, Inc.||Use of a Composition which Regulates Oxidation/Reduction Reactions Intracellularly and/or Extracellularly in a Staining or Sorting Process|
|US20080124805 *||4 May 2007||29 May 2008||Honeywell International Inc.||Cytometer having fluid core stream position control|
|US20090287421 *||27 Jul 2005||19 Nov 2009||George C Malachowski||Enhancing Flow Cytometry Discrimination with Geometric Transformation|
|US20130081446 *||4 Apr 2013||Azbil Corporation||Particle detecting device evaluating system and particle detecting device evaluating method|
|USRE29141 *||3 Dec 1975||22 Feb 1977||Coulter Electronics, Inc.||Apparatus for orienting generally flat particles for sensing|
|DE3219275A1 *||21 May 1982||16 Dec 1982||Technicon Instr||Verfahren und geraet zum korrigieren von koinzidenzfehlern beim erfassen und zaehlen von miteinander gemischten dominanten und nicht dominanten teilchen|
|EP0046345A2 *||28 Jul 1981||24 Feb 1982||Ortho Diagnostic Systems Inc.||Controlled hydrodynamic flow in flow cytometry systems|
|EP0101161A2 *||20 Jun 1983||22 Feb 1984||TECHNICON INSTRUMENTS CORPORATION (a New York corporation)||Apparatus and method for passing two fluids simultaneously through an analytical flow cell|
|EP0107333A2 *||19 Sep 1983||2 May 1984||TECHNICON INSTRUMENTS CORPORATION (a New York corporation)||Apparatus and method for supply of sample and sheath liquids to analytical flow cell|
|EP0118896A1 *||9 Mar 1984||19 Sep 1984||E.I. Du Pont De Nemours And Company||Particle counting apparatus|
|EP0195488A2 *||19 Mar 1986||24 Sep 1986||Philips Electronics N.V.||Spectrometer|
|EP0347696A2 *||10 Jun 1989||27 Dec 1989||BASF Aktiengesellschaft||Apparatus for measuring the rate of dispersion in flowing suspensions|
|EP0360487A2 *||13 Sep 1989||28 Mar 1990||Hitachi, Ltd.||Method and apparatus for analysis of particles contained in a liquid sample|
|EP0545284A1 *||26 Nov 1992||9 Jun 1993||Canon Kabushiki Kaisha||Sample measuring device and sample measuring system|
|EP1348943A2 *||24 Mar 2003||1 Oct 2003||Sysmex Corporation||Sheath liquid for particle analyzer|
|WO1980002198A1 *||10 Apr 1980||16 Oct 1980||Leitz Ernst Gmbh||Device for hydrodynamic focussing of a particle-suspension in a liquid flow cytophotometer|
|U.S. Classification||356/36, 356/246, 356/335, 250/576, 356/442, 250/222.2|
|International Classification||G01N21/03, G01N21/05, G01N15/14|
|Cooperative Classification||G01N21/05, G01N15/1404|
|European Classification||G01N15/14C, G01N21/05|
|5 Jul 1988||AS||Assignment|
Owner name: TECHNICON INSTRUMENTS CORPORATION
Free format text: MERGER;ASSIGNOR:REVGROUP PANTRY MIRROR CORP.;REEL/FRAME:004912/0740
Effective date: 19871231