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WO1985005680A1 - Systemes optiques pour cytometres a ecoulement - Google Patents

Systemes optiques pour cytometres a ecoulement Download PDF

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Publication number
WO1985005680A1
WO1985005680A1 PCT/US1985/000989 US8500989W WO8505680A1 WO 1985005680 A1 WO1985005680 A1 WO 1985005680A1 US 8500989 W US8500989 W US 8500989W WO 8505680 A1 WO8505680 A1 WO 8505680A1
Authority
WO
WIPO (PCT)
Prior art keywords
waveguides
aperture
optical
orifice plate
flow
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.)
Ceased
Application number
PCT/US1985/000989
Other languages
English (en)
Inventor
Howard M. Shapiro
Michael Hercher
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.)
Individual
Original Assignee
Individual
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
Application filed by Individual filed Critical Individual
Publication of WO1985005680A1 publication Critical patent/WO1985005680A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1434—Optical arrangements
    • G01N15/1436—Optical arrangements the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1434—Optical arrangements
    • G01N2015/144—Imaging characterised by its optical setup
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1486—Counting the particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/08—Optical fibres; light guides

Definitions

  • This invention pertains to the flow cytometers and more particularly to such devices wherein a number of different optical measurements are simultaneously made.
  • Optical flow cytometers in which cells having predetermined optical characteristics are detected, counted, and/or sorted as they pass an observing station, are well known.
  • the individual cells are illuminated and observed, preferably simultaneously, by one or more optical channels. Observations may be made over one or more wavelength bands and at a variety of angles to the incident illumination. Cells are counted or sorted according to the observed values of the the appropriate optical properties (e.g., color, scattering characteristics, fluorescent signature, optical density, or the like) .
  • the appropriate optical properties e.g., color, scattering characteristics, fluorescent signature, optical density, or the like.
  • a variety of parameters are observed and the cells are classified for counting or sorting according to the correlation of these various parameters.
  • lenses usually microscope objectives
  • a typical multi-parameter "'"' flow cytophotometer incorporates a number of high-quality microscope objectives, together with their precision alignment mechanisms, impacting costs of parts, assembly, and routine maintenance.
  • an object of the present invention is to provide an optical apparatus for use in flow cytometers wherein a maximum number of optical channels may be situated in one plane transverse to the flow stream.
  • Another object of the present invention is to provide an optical apparatus for use in flow cytometers that permits close spacing of a number of observing stations, or observing stations and ancillary systems, along the flow stream.
  • a further object of the present invention is to provide an optical apparatus for use in flow cytometers that insures alignment of a number of associated optical paths without the requirement for time-consuming alignment procedures or costly precision alignment devices.
  • Yet another object of the present invention is to provide an optical system for multi-parameter flow cytophotometers which is relatively inexpensive.
  • an orifice plate used to define the flow stream, serves as a substrate to support a plurality of optical waveguides.
  • the waveguides terminate at the aperture in the orifice plate through which the flow stream passes.
  • the ends of the waveguides distal from the aperture are variously coupled to illuminatio sources and detectors.
  • the waveguides are optical fibers bonded to the orifice plate.
  • the orifice plate serves as the substrate for an integrated optics device, the waveguides being provided by appropriate implantation.
  • the orfice plate serves as the mechanical mounting for the waveguides, and the waveguides are bonded thereto, the mechanical constraints of normal optical-mechanical structures are avoided. Consequently, a maximum number of coplanar optical channels radiating from a single center can be provided. Further, as the alignment is built into the system, elaborate precision alignment devices are not required, nor is there any need for time-consuming alignment procedures. It will also be recognized that the aperture plate and waveguide structure of the present invention may be fabricated by relatively straight forward procedures, resulting in a relatively inexpensive optical system.
  • FIG. 1 is a perspective view of a preferred embodiment of the present invention, as seen from an angle oblique to the direction of the flow stream of a flow cytometer;
  • Fig. 2 is a fragmentary plan view of the embodiment of Fig. 1, viewed parallel to the direction of the flow stream;
  • Fig. 3 is a fragmentary sectional view taken along the lines 3-3 of Fig. 2, showing the embodiment of Fig. 1 and the immediately adjacent structure of a flow cytometer.
  • FIG. 1 there may be seen a a preferred embodiment of the present invention.
  • the fluid stream of a flow cytometer (not shown) is made to flow past one or more observing stations 10, as by gravity or pump means well known in the art.
  • Each observing station 10 comprises an orifice plate 12, preferably in the form of a thin, flat plate provided with a small, centrally located, circular aperture 14, and a plurality of optical waveguides 16.
  • Orifice plate 12 is installed in a cytometer so that the plane of the plate is transverse to the direction of flow of the fluid stream, which, it will be understood, is directed at the plate so as to pass through aperture 14, as schematically indicated by arrow 17.
  • orifice plate 12 is preferably of a material chosen to be impermeable to and non-reactive with the fluids of interest.
  • Typical materials of construction of orifice plate 12 include, but are not limited to, glass, fused silica, saphire, polypropylene, polyolefin, nylon, or the like. It will be understood that, if necessary, orifice plate 12 may be supplied with a coating to provide physical isolation between it and the flow stream or optical isolation between it and waveguides 16, if necessary.
  • the overall dimensions of orifice plate 12 are not particularly critical, and are chosen as convenient. Typical dimensions are a few centimeters square by one millimeter thick, although other dimensions may be accommodated.
  • aperture 14 is a substantially right circular cylindrical bore, substantially normal to and penetrating through orifice plate 12.
  • the bore forming the aperture may be a right regular prism or a right circular conic frustum. In the latter case, the aperture will normally be placed in the flow stream with the smaller diameter base of the frustum upstream of the larger.
  • the dimensions of aperture 14 are set primarily by considerations of the cells to be observed, and for most cases will be typically between about 20 micrometers and about 200 micrometers in diameter.
  • aperture 14 is dimensioned to permit free, undisturbed passage of cells flowing substantially along the axis of the aperture, and to dispose the cells a sufficient distance from the walls of the aperture to permit adequate illumination and viewing, as will become apparent.
  • Aperture 14 may be formed in orifice plate 12 by a variety of means, such as micromachining, chemical milling, laser drilling, or the like. As already noted, aperture 14 is preferably centrally situated in orifice plate 12.
  • Optical waveguides 16 may take the form of individual optical fibers attached to orifice plate 12, or may be portions of an integrated optical device implanted or otherwise formed on or in the orifice plate, as by casting, ion implantation, vapor deposition, or the like.
  • a preferred embodiment employs cladded optical fibers.
  • waveguides 16 are arranged substantially in a single plane (as, for instance, a surface of orifice plate 12) so as to extend radially from aperture 14. An end of each waveguide terminates at aperture 14 such that the optical axis of the waveguide is substantially normal to the axis of the aperture (and the flow stream) .
  • up to six optical fibers each 100 micrometers in diameter, may be so disposed about a 200 micrometer diameter aperture (FIG. 2) ; obviously, a larger number of waveguides of smaller diameter could be disposed about the same size aperture.
  • a particularly convenient method for insuring that the individual waveguides terminate at aperture 14 is to initially fabricate the waveguide and orifice plate structure without an aperture. Aperture 14 is then drilled through the center of an abutted fan of waveguides, as by laser drilling. This approach is particularly suited to the construction of a pair of waveguides intended for transmission measurements: drilling through a single straight waveguide assures the desired 180 degree alignment of the resulting pair of waveguides.
  • the ends of the waveguides distal from aperture 14. are coupled to various optical sources and detectors, schematically represented by index numbers 18, in accordance with the measurements to be undertaken, as will be understood by those skilled in the art.
  • Waveguides 16 are variously used for illumination and collection, and it will be understood that the form of the waveguide may be varied accordingly.
  • single-mode optical fibers having graded index sheaths may be used as desired for illumination, in order to provide a narrow illuminating beam.
  • Commercially available fibers having an outer diameter of about 100 micrometers may be used with visible laser sources to provide a beam having a beam diameter at the fiber face of about 5 micrometers, spreading to some 70 micrometers as it passes across a 200 micrometer orifice.
  • multi-mode fibers may be advantageous because of their larger optical throughput.
  • waveguides 16 The exact configuration of waveguides 16 depends on the optical parameters to be observed, as is well known in the art of cytophotometry. Thus, transmission measurements require illuminating and collecting waveguides to be in line with one another. Fluorescent measurements may be made with illuminating and collecting waveguides not in line, the difference in viewing angle providing some isolation of the illuminating and fluorescent radiation. The observation of scattering signatures requires collection at a number of angles to the illumination direction.
  • the axes of waveguides 16 preferably should be disposed substantially radially to aperture 14 proximate to the aperture, in order to illuminate and view the core of the flow stream passing through the aperture.
  • the radial orientation of the waveguides may be maintained distal from the aperture, and the waveguides may be curved, as necessary, in order to more conveniently accommodate sources and detectors.
  • the optical system of the present invention has a number of advantages.
  • the orfice plate serves as the mechanical mounting for the waveguides, and the waveguides are bonded thereto, the mechanical constraints of normal optical-mechanical structures are avoided. Consequently, a maximum number of coplanar optical channels radiating from a single center can be provided.
  • the alignment is built into the system, elaborate precision alignment devices are not required, nor is there any need for time-consuming alignment procedures.
  • the aperture plate and waveguide structure of the present invention may be fabricated by relatively straightforward procedures, resulting in a relatively inexpensive optical system.
  • waveguides 16 might be other optical devices than cladded fibers.
  • waveguides 16 might be integrated optical waveguides implanted in or on orifice plate 12 as by ion beam implantation, vapor deposition, or the like.
  • fiber waveguides may be recessed into the surface of orifice plate 12, rather than attached to the surface.
  • the use of cladded fibers cemented to the surface merely allows ready assembly of individual custom-made observing stations 10, since the various masks and processing steps normally required for the production of integrated optical devices are not required. Fibers may be simply cemented, as with epoxy adhesive, to orifice plate 12 in the desired configuration.
  • cladded fibers make it unnecessary to otherwise optically isolate either the orifice plate from the waveguides or closely adjacent waveguides from one another. It will be appreciated, however, that uncladded fibers might be used, in which case the orifice plate would have to be optically isolated from the waveguides, as by a suitable coating or an appropriate choice of materials, and the waveguides either spaced well apart or otherwise isolated from one another in order to prevent cross-talk.
  • An integrated-optic observing station similar to that described for the case of a fiber optic station, is a preferred embodiment for quantity production of observing stations, as such devices .may be quantity produced relatively cheaply once the initial tooling is available.
  • the full scope of integrated optics may be used to provide additional optical elements, such as filters, dispersive devices, and the like, on the substrate provided by orifice plate 12.
  • Such an approach for the fabrication of observing stations also offers the possibility of integrating sources (such as LEDs) and detectors (such as photodiodes) , and electronics into the manufacturing process, building, in effect, an integrated chip containing source, optics, detectors, and electronics.
  • waveguides 16 may be similarly disposed about aperture 14 on both surfaces of an orifice plate 12, as shown in FIG. 3. In this way, a pair of closely spaced embodving stations may be disposed along the flow stream.
  • FIG. 3 also indicates the manner in which one or more observing stations 10 are positioned in the flow stream of a cytometer.
  • the cells to be observed are entrained in a core flow stream 20 that is centered in a sheath fluid 22 conducted through a channel 24.
  • Core stream 20 is injected into the center of channel 24 by an injection channel 26 that terminates a short distance up stream from the observing station and in line with aperture 14.
  • Core stream 20 necks down as it passes through aperture 14, and the cells entrained in the core flow are directed substantially centrally through the aperture.
  • a plurality of aperture plates may be disposed in closely spaced arrangement along the direction of flow of the fluid stream.
  • orifice plate 12 of observation station 10 may also serve as the substrate for other components of a flow cytometer.
  • the surfaces of the orifice plate or of aperture 14 might be made electrically conductive, to serve as an electrical counting or measuring device or the charging electrode or ground ring of a electrostatic cell sorting mechanism.
  • the orifice plate may form the substrate of an acoustic transducer to serve as the droplet generator of a sorting mechanism.
  • optical system of the present invention may be used to observe particulates other than cells in a flow stream.

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  • Chemical & Material Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Optical Measuring Cells (AREA)

Abstract

Un système optique (10) utilisé dans des cytomètres à écoulement comprend une pluralité de guides d'ondes (16) maintenus sur une plaque d'orifice (12) utilisée pour définir le courant d'écoulement (17). Les guides d'ondes (16) se terminent à une ouverture (14) dans la plaque d'orifice (12) à travers laquelle on fait passer le courant d'écoulement (17). Les extrémités des guides d'ondes (16) éloignées de l'ouvertue (14) sont couplées de différentes façons aux sources d'éclairage (18) et au détecteur (18). Dans un mode préféré de réalisation, les guides d'ondes (16) sont des fibres optiques soudées à la plaque d'orifice (12). Dans une variante, la plaque d'orifice (12) sert de substrat pour un dispositif optique intégré, le guide d'ondes (16) étant réalisé grâce à une implantation appropriée.
PCT/US1985/000989 1984-06-01 1985-05-28 Systemes optiques pour cytometres a ecoulement Ceased WO1985005680A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US61647984A 1984-06-01 1984-06-01
US616,479 1984-06-01

Publications (1)

Publication Number Publication Date
WO1985005680A1 true WO1985005680A1 (fr) 1985-12-19

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EP (1) EP0183798A1 (fr)
WO (1) WO1985005680A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003008937A3 (fr) * 2001-07-18 2003-08-21 Univ Michigan Cytometres de flux et systeme de detection d'une taille reduite
EP1403632A1 (fr) * 2002-09-27 2004-03-31 Becton, Dickinson and Company Optique d'excitation à base de prismes pour cytométrie à flux
WO2005085803A1 (fr) * 2004-03-01 2005-09-15 Firma Cytecs Gmbh Dispositif permettant de mesurer la lumiere emise par des cellules biologiques ou particules microscopiques
FR2878032A1 (fr) * 2004-11-18 2006-05-19 Horiba Abx Sa Sa Dispositif d'inspection d'un fluide par illumination uniforme au moyen d'un guide de lumiere conforme
US7443491B2 (en) * 2002-12-03 2008-10-28 Bay Bioscience Kabushiki Kaisha System for collecting information on biological particles
WO2010099118A1 (fr) * 2009-02-27 2010-09-02 Beckman Coulter, Inc. Système optique stabilisé pour cytométrie de flux
US20120196314A1 (en) * 2007-09-10 2012-08-02 The Penn State Research Foundation Three-dimensional (3d) hydrodynamic focusing using a microfluidic device
WO2016001324A1 (fr) * 2014-07-01 2016-01-07 Octrolix Bv Système et procédé de cytométrie en flux
EP1739402B1 (fr) * 2004-04-23 2017-08-02 The Furukawa Electric Co., Ltd. Procedes de separation, d"identification et d'administration de specimen et dispositif correspondant, et procede de dispositif d"analyse
JP2017532545A (ja) * 2014-09-09 2017-11-02 ヘレーウス クヴァルツグラース ゲゼルシャフト ミット ベシュレンクテル ハフツング ウント コンパニー コマンディートゲゼルシャフトＨｅｒａｅｕｓ Ｑｕａｒｚｇｌａｓ ＧｍｂＨ ＆ Ｃｏ． ＫＧ ファイバコアの中心に沿って、試料を収容するための中空管路を備えている光ファイバ

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Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7105355B2 (en) 2001-07-18 2006-09-12 The Regents Of The University Of Michigan Flow cytometers and detection system of lesser size
WO2003008937A3 (fr) * 2001-07-18 2003-08-21 Univ Michigan Cytometres de flux et systeme de detection d'une taille reduite
US7381565B2 (en) 2001-07-18 2008-06-03 The Regents Of The University Of Michigan Flow cytometers and detection system of lesser size
EP1403632A1 (fr) * 2002-09-27 2004-03-31 Becton, Dickinson and Company Optique d'excitation à base de prismes pour cytométrie à flux
EP1574838A4 (fr) * 2002-12-03 2011-09-28 Bay Bioscience Kabushiki Kaisha Dispositif permettant la collecte d'informations sur une particule biologique
US7443491B2 (en) * 2002-12-03 2008-10-28 Bay Bioscience Kabushiki Kaisha System for collecting information on biological particles
WO2005085803A1 (fr) * 2004-03-01 2005-09-15 Firma Cytecs Gmbh Dispositif permettant de mesurer la lumiere emise par des cellules biologiques ou particules microscopiques
EP1739402B1 (fr) * 2004-04-23 2017-08-02 The Furukawa Electric Co., Ltd. Procedes de separation, d"identification et d'administration de specimen et dispositif correspondant, et procede de dispositif d"analyse
FR2878032A1 (fr) * 2004-11-18 2006-05-19 Horiba Abx Sa Sa Dispositif d'inspection d'un fluide par illumination uniforme au moyen d'un guide de lumiere conforme
US7724371B2 (en) 2004-11-18 2010-05-25 Horiba Abx Sas Device for examining a fluid by uniform illumination using a configured light guide
WO2006053960A1 (fr) * 2004-11-18 2006-05-26 Horiba Abx Sas Dispositif d'inspection d'un fluide par illumination uniforme au moyen d'un guide de lumiere conforme
US20120196314A1 (en) * 2007-09-10 2012-08-02 The Penn State Research Foundation Three-dimensional (3d) hydrodynamic focusing using a microfluidic device
US8941826B2 (en) * 2007-09-10 2015-01-27 The Penn State Research Foundation Three-dimensional (3D) hydrodynamic focusing using a microfluidic device
WO2010099118A1 (fr) * 2009-02-27 2010-09-02 Beckman Coulter, Inc. Système optique stabilisé pour cytométrie de flux
US9453791B2 (en) 2014-07-01 2016-09-27 Octrolix Bv Flow cytometry system and method
KR20170023176A (ko) * 2014-07-01 2017-03-02 옥트로릭스 비브이 유동 세포 계측 시스템 및 방법
CN106662520A (zh) * 2014-07-01 2017-05-10 奥克特罗利克斯有限责任公司 流式细胞术系统和方法
WO2016001324A1 (fr) * 2014-07-01 2016-01-07 Octrolix Bv Système et procédé de cytométrie en flux
US10180390B2 (en) 2014-07-01 2019-01-15 Lionix International Bv Flow cytometry system and method
KR102217008B1 (ko) 2014-07-01 2021-02-18 옥트로릭스 비브이 유동 세포 계측 시스템 및 방법
JP2017532545A (ja) * 2014-09-09 2017-11-02 ヘレーウス クヴァルツグラース ゲゼルシャフト ミット ベシュレンクテル ハフツング ウント コンパニー コマンディートゲゼルシャフトＨｅｒａｅｕｓ Ｑｕａｒｚｇｌａｓ ＧｍｂＨ ＆ Ｃｏ． ＫＧ ファイバコアの中心に沿って、試料を収容するための中空管路を備えている光ファイバ

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Publication number Publication date
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