EP2572182A2 - Agencement pour mesurer les propriétés optiques de particules dans une dispersion - Google Patents

Agencement pour mesurer les propriétés optiques de particules dans une dispersion

Info

Publication number
EP2572182A2
EP2572182A2 EP11782374A EP11782374A EP2572182A2 EP 2572182 A2 EP2572182 A2 EP 2572182A2 EP 11782374 A EP11782374 A EP 11782374A EP 11782374 A EP11782374 A EP 11782374A EP 2572182 A2 EP2572182 A2 EP 2572182A2
Authority
EP
European Patent Office
Prior art keywords
arrangement according
laser
measuring
light
cuvette
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.)
Withdrawn
Application number
EP11782374A
Other languages
German (de)
English (en)
Inventor
Wolfgang GÖHDE
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.)
Partec GmbH
Original Assignee
Partec GmbH
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 Partec GmbH filed Critical Partec GmbH
Publication of EP2572182A2 publication Critical patent/EP2572182A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N21/0303—Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • 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
    • G01N2015/1006—Investigating individual particles for cytology
    • 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
    • G01N2201/06—Illumination; Optics
    • G01N2201/061—Sources
    • G01N2201/06113—Coherent sources; lasers

Definitions

  • the task is in addition to the rapid detection of the individual particles is to measure them as accurately as possible optically.
  • lasers are used as light sources for the forward and side scattered light measurements as well as for the fluorescence excitation.
  • the laser light is allowed to act on the particles with a very small opening angle (low divergence) or as a parallel light beam.
  • the metrological problem consists in the fact that morphologically complex particles, which can often be structured like very flat, flat objects, are achieved by the laser beam at right angles to the particle surface or parallel to the particle surface. This leads, for example, to "correct" measured values of the particle fluorescence in the case of planar illumination and to "erroneous" measured values when the laser light hits the edge of the particle.
  • the invention relates to an arrangement which avoids this measurement error.
  • the flow cytometric analysis of microscopic particles has great economic importance if it is possible to make the measurements highly precise.
  • One of these Applications is the measurement of the DNA content of sperm. If this measurement is accurate enough, the 2 varieties of sperm containing X and Y chromosomes can be sorted with a device downstream of the flow cytometer and used for animal breeding.
  • the object of this invention is to find a way that in the previously known Laser-based flow cytometers reduce or completely eliminate the erroneous influence of shape factors on the measurement accuracy. This leads to a significantly increased measurement accuracy.
  • the object of the invention is also to combine the high measurement accuracy achieved with the two scattered light parameters. These can not be measured with conventional light sources.
  • a device for laser light excitation is proposed, which avoids the influence of form factors. Compared to conventional light sources with the advantage of high numerical aperture excitation, laser excitation allows a much higher density of light energy, resulting in more accurate measurements (better signal-to-noise ratio).
  • FIG. 2 is a plan view of a first embodiment of a flow cytometer
  • Fig. 5 is a plan view of a third embodiment of a flow cytometer.
  • Fig. 6 is a plan view of a fourth embodiment of a flow cytometer.
  • Fig. 1 shows in the manner of a schematic vertical section the structure of the Bruvettenteils a flow cytometer and the arrangement of the excitation beam path with the aid of a laser 1 and the measuring beam path with light-collecting optics in shape a converging lens 5, a laser light blocking filter 6 and the photodetector 7.
  • the dispersion of the measuring cuvette 3 is fed through a narrow tube 21.
  • Particle-free medium 14 is fed to the particle flow, which leads to a centering of the particles when passing the measuring range at 23.
  • the morphologically different particles have no preferential orientation. From FIGS. 2 and 4 to 6, the measuring cuvette 3 is in each case in the
  • the laser light excitation takes place from two directions, wherein the irradiation direction of the laser light beam 1 is offset from the laser light beam 2 by 90 °.
  • the laser light in the form of the two laser light beams 1 and 2 excites the fluorescence from two sides of the cuvette 3.
  • a second light collecting optic or converging lens 9 is mounted, which is offset from the first condenser lens 8 by 90 °. This type of detection of the light emanating from each particle further reduces the influence of form factors. Both beam paths for light detection are with high numerical aperture optics in
  • Fig. 3 shows an epithelial cell, which may have up to 60 ⁇ diameter in humans. Will this be on the edge? lights, the laser light reaches the nucleus 16 only weakened. If this cell is irradiated flat and angular at the same time, the jetting errors cancel each other out.
  • the substance to be measured is located in the sperm head 17, in the middle part 18 is preferably RNA. If such an object is irradiated edgewise, the laser light does not reach all the DNA components uniformly, some of the excitation light is "scattered away", and within the sperm head 17 it absorbs light, with the result that not all DNA components contribute equally Fluorescence be excited.
  • FIG. 4 shows the arrangement with laser double excitation in a spatial representation.
  • a measuring optics with a converging lens 5 a laser light blocking filter 6 and a photodetector 7 is shown.
  • the condenser lens 5 has a high numerical aperture, so that the influence of form factors is avoided on the measurement side. Deviating from this illustrated embodiment can be provided, as in the embodiment of FIG. 2, two laser light beams 1 and 2 each assign their own measuring optics.
  • FIG. 5 shows a further embodiment of the measuring apparatus according to the invention: Since the available laser light blocking filters 6, 10, 11 do not completely block the exciting laser light at some wavelengths, an arrangement is provided in FIG. 5 which prevents the respective Laser light beam 1, 2 can go directly into the measuring optics.
  • the optical axis of the converging lens 5 with respect to the beam directions of the two excitation laser light beams 1, 2 arranged in the bisector, ie at an angle of 45 ° or 135 °.
  • a special shape of the Measuring cuvette 19 with a designated as an inclined surface 20 fifth surface in cross section is provided. This fifth surface allows the attachment of a measuring optics with high numerical aperture, without laser excitation light enters the measuring beam path.
  • the parallel alignment of the front surface of the converging lens 5 to the inclined surface 20 ensures that the fluorescent light from the measuring cuvette 19 into the converging lens 5 with low loss.
  • FIG. 6 shows a measuring cuvette which is designed essentially like that of FIG. 5 and is therefore likewise designated by 19. Due to their two aligned at an angle of 90 ° to each other, adjacent to the inclined surface 20 surfaces in addition to the converging lens 5, which is associated with the inclined surface 20, the arrangement of two converging lenses 8, 9 are provided at an angle of 90 ° to each other, similar this is shown in Fig. 2. With this arrangement, particularly precise measurement results can be achieved:
  • Notch filters 21 and 22 are provided between the measuring cuvette 19 and the converging lenses 8 and 9. They block the unwanted light components and are only permeable to stray light if possible. Since only scattered light components are to be detected, ie light components that are incident at an angle of more than 0 °, the directly incident laser light is blocked with so-called laser stops 24.
  • the alignment of the cell within the measuring cuvette 19 can be calculated so that, for example, the measurement results of cells that are unfavorably aligned are not sufficient for further investigation be taken into account, or so that the measurement results of the fluorescent light certain correction factors - depending on the orientation of the measured cell - can be assigned.
  • the illustrated embodiments may be provided to direct the light from the measuring cuvette in the direction of the photodetector by an optical element which is designed as a cylinder with a cylindrical reflection surface.
  • the cylinder may be designed as a hollow cylinder whose inner surface forms the cylindrical reflection surface, or is designed as a solid, translucent cylinder whose outer surface forms the cylindrical reflection surface.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Dispersion Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biophysics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Optical Measuring Cells (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

L'invention concerne un agencement pour mesurer les propriétés optiques de particules dans une dispersion fluide au moyen d'une cuvette de mesure, dont l'espace intérieur central est traversé par la dispersion et par la lumière d'un rayon laser. Selon l'invention, l'espace intérieur de la cuvette est éclairé par deux rayons laser qui forment un angle de 90° l'un par rapport à l'autre.
EP11782374A 2010-05-18 2011-05-13 Agencement pour mesurer les propriétés optiques de particules dans une dispersion Withdrawn EP2572182A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102010017015 2010-05-18
PCT/DE2011/075110 WO2011144208A2 (fr) 2010-05-18 2011-05-13 Agencement pour mesurer les propriétés optiques de particules dans une dispersion

Publications (1)

Publication Number Publication Date
EP2572182A2 true EP2572182A2 (fr) 2013-03-27

Family

ID=44970902

Family Applications (1)

Application Number Title Priority Date Filing Date
EP11782374A Withdrawn EP2572182A2 (fr) 2010-05-18 2011-05-13 Agencement pour mesurer les propriétés optiques de particules dans une dispersion

Country Status (6)

Country Link
US (2) US20130070243A1 (fr)
EP (1) EP2572182A2 (fr)
JP (1) JP2013526714A (fr)
CA (1) CA2784073A1 (fr)
WO (1) WO2011144208A2 (fr)
ZA (1) ZA201204092B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8624850B2 (en) 2007-03-14 2014-01-07 Power2B Displays and information input devices

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USD673286S1 (en) * 2010-04-29 2012-12-25 Sony Corporation Micro flow channel chip
USD673287S1 (en) 2010-11-24 2012-12-25 Sony Corporation Micro flow channel chip
USD869308S1 (en) 2010-04-29 2019-12-10 Sony Corporation Micro flow channel chip
US9746412B2 (en) 2012-05-30 2017-08-29 Iris International, Inc. Flow cytometer

Citations (2)

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US4999513A (en) * 1988-09-09 1991-03-12 Canon Kabushiki Kaisha Particle measuring apparatus
DE102005055825A1 (de) * 2005-11-23 2007-05-24 Basf Ag Vorrichtung und Verfahren für die automatische Bestimmung der individuellen dreidimensionalen Partikelform

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US3819270A (en) * 1972-10-02 1974-06-25 Block Engineering Blood cell analyzer
JPS62168033A (ja) * 1986-01-18 1987-07-24 Canon Inc 粒子解析装置
EP0279000B1 (fr) * 1987-02-17 1993-07-21 Ratcom, Inc. Cytométrie à écoulement
US5041733A (en) * 1987-03-20 1991-08-20 Agency Of Industrial Science & Technology Method and apparatus for identifying chromosomes or cells
JPH0641918B2 (ja) * 1987-03-27 1994-06-01 工業技術院長 流れる染色体または細胞に照射するレ−ザ光の照射方法およびその装置
JPH02138851A (ja) * 1988-11-18 1990-05-28 Canon Inc 粒子測定装置
JPH0621859B2 (ja) * 1988-09-09 1994-03-23 キヤノン株式会社 粒子測定装置
US5760900A (en) * 1989-03-18 1998-06-02 Canon Kabushiki Kaisha Method and apparatus for optically measuring specimen
JP2611015B2 (ja) * 1989-11-30 1997-05-21 ホーヤ・ショット株式会社 移動物体の表面反射光測定装置
JP2899368B2 (ja) * 1990-04-09 1999-06-02 興和株式会社 粒子測定装置
JP3145487B2 (ja) * 1992-06-12 2001-03-12 シスメックス株式会社 粒子分析装置
US5489977A (en) * 1993-08-11 1996-02-06 Texaco Inc. Photomeric means for monitoring solids and fluorescent material in waste water using a falling stream water sampler
JP3640461B2 (ja) * 1996-04-03 2005-04-20 シスメックス株式会社 粒子分析装置
US6139800A (en) * 1997-06-23 2000-10-31 Luminex Corporation Interlaced lasers for multiple fluorescence measurement
WO2002031182A2 (fr) * 2000-10-12 2002-04-18 Amnis Corporation Procede et appareil de lecture de billes marquees par rapporteurs
GB0307756D0 (en) * 2003-04-03 2003-05-07 Suisse Electronique Microtech Measuring the concentration and motility of light scattering particles
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GB0513128D0 (en) * 2005-06-27 2005-08-03 Ojk Consulting Ltd An optical arrangement for a flow cytometer
US8031340B2 (en) * 2006-02-22 2011-10-04 Accuri Cytometers, Inc. Optical system for a flow cytometer
JP4990746B2 (ja) * 2007-12-14 2012-08-01 ベイバイオサイエンス株式会社 液体フローに含まれる生物学的粒子を分別する装置ならびにその方法

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4999513A (en) * 1988-09-09 1991-03-12 Canon Kabushiki Kaisha Particle measuring apparatus
DE102005055825A1 (de) * 2005-11-23 2007-05-24 Basf Ag Vorrichtung und Verfahren für die automatische Bestimmung der individuellen dreidimensionalen Partikelform

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO2011144208A2 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8624850B2 (en) 2007-03-14 2014-01-07 Power2B Displays and information input devices

Also Published As

Publication number Publication date
US20140374623A1 (en) 2014-12-25
JP2013526714A (ja) 2013-06-24
ZA201204092B (en) 2013-06-26
CA2784073A1 (fr) 2011-05-24
WO2011144208A3 (fr) 2012-04-12
US20130070243A1 (en) 2013-03-21
WO2011144208A2 (fr) 2011-11-24

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