WO2001077645A1 - Equipement d'eclairage et de reproduction, notamment lors de la realisation de tests d'immunofluorescence - Google Patents

Equipement d'eclairage et de reproduction, notamment lors de la realisation de tests d'immunofluorescence Download PDF

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Publication number
WO2001077645A1
WO2001077645A1 PCT/DE2001/001301 DE0101301W WO0177645A1 WO 2001077645 A1 WO2001077645 A1 WO 2001077645A1 DE 0101301 W DE0101301 W DE 0101301W WO 0177645 A1 WO0177645 A1 WO 0177645A1
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WO
WIPO (PCT)
Prior art keywords
medium
receiver
medium surfaces
carrier
light
Prior art date
Application number
PCT/DE2001/001301
Other languages
German (de)
English (en)
Inventor
Peter Boll
Dieter Krause
Franz Drobner
Christian Kassel
Original Assignee
Siemens Production And Logistics Systems Ag
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 Siemens Production And Logistics Systems Ag filed Critical Siemens Production And Logistics Systems Ag
Publication of WO2001077645A1 publication Critical patent/WO2001077645A1/fr

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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
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Definitions

  • the present invention relates to a device for illumination and imaging, in particular when quantitative fluorescence immunoassays are carried out, according to the preamble of claim 1.
  • a medium carrier which consists of an upper part, a lower part and an intermediate part arranged between them. Antibody strips or medium surfaces running in the transverse direction and spaced apart in the longitudinal direction are applied to the side of the lower part facing the upper part.
  • the intermediate layer has a recess which runs in the longitudinal direction and leaves all medium surfaces free.
  • two through holes spaced apart in the longitudinal direction are arranged, one of which forms an access to the recess of the intermediate part on one side of the medium surfaces and the other forms an access to the recess of the intermediate part on the other side of the medium surfaces.
  • Containers are pumped out and vice versa, whereby the medium surfaces react with the sample liquid.
  • the analytically specific Antibodies on the medium surfaces are marked with a marking substance which is optically excited at a substance-specific wavelength and which detects the fluorescent light, which has a different wavelength, which is generally greater than the wavelength of the excitation light, with a suitable detector while evaluating the fluorescence intensity , It is known to illuminate the entire surface of the medium with laser light and to supply the fluorescent light via optics to a specially designed receiver for the fluorescent light.
  • the object of the present invention is to improve a device for illumination and imaging, in particular when carrying out fluorescence immunoassays, in such a way that a high imaging constancy can be achieved.
  • the main advantage of the present invention is that a high imaging constancy is obtained as a result of the selective scanning of the individual medium areas. This also applies to high excitation intensity.
  • Another advantage of the present invention is that the optics used for imaging can be relatively small, because only the fluorescent light emitted by a medium surface has to be detected and imaged on a receiver.
  • Another advantage of the present invention is that the partial lighting fades the Medium areas are largely avoided by illuminating only individual medium areas.
  • the individual medium surfaces are advantageously each exposed to the same irradiation time.
  • the background fluorescence of the medium carrier is advantageously reduced by the partial illumination.
  • FIG. 1 shows a schematic representation of the structure of a medium carrier
  • FIG. 2 shows a schematic illustration of the structure of the device for illumination and imaging according to the invention, in particular in the case of quantitative fluorescence immunoassays;
  • FIG. 3 shows a further embodiment of the invention.
  • the upper part of a medium carrier 1 is designated 3.
  • the reference numeral 5 denotes the intermediate part and the reference numeral 7 denotes the lower part of the medium carrier 1.
  • the lower part 7 consists of a transparent, high-index material, such as glass or plastic.
  • the intermediate part 5 preferably has the shape of a thin film which is provided on both sides with an adhesive film.
  • the upper part 3 has a refractive index which is smaller than that of the lower part 7.
  • the intermediate part 5 whose thickness m is of the order of magnitude of approximately 50 ⁇ m, there is a recess 11, which runs in the longitudinal direction of the intermediate part 5, and is dimensioned such that when the intermediate part 5 is fastened to the lower part 7, in particular it sticks is, the medium surfaces 9 m of the recess 11 are.
  • the upper part 3 has through holes 13 and 15 spaced apart from one another in the longitudinal direction, of which the through hole 13 connects to the recess on one side of the medium surfaces 9 and the other through hole 15 connects to the recess 11 on the other side of the Medium surfaces 9 m produces the material carrier 1.
  • Material strip carriers 1 of this type are known.
  • the present device for illuminating and imaging such a medium carrier 1, the present device according to FIG. 2 essentially comprises a light source, preferably a laser source 17, an imaging optics 19, which are arranged in a common housing body 21, which in relation to the medium carrier 1 in the longitudinal direction thereof, ie m in the direction of the row of medium surfaces, can be moved, as shown by arrow 23.
  • the housing body 21 is moved with respect to the medium carrier 1 by a spmdelant ⁇ eb, which comprises a stepping motor 25 and a spindle 27, which runs through nut parts 29 of the housing body 21, around this along the travel path indicated by the arrow 23 in a sled-like manner to move.
  • the housing body 21 engages a guide (not shown) on its side opposite the spindle 27 with a support 47 m. It is pointed out that other mechanisms or drives are also conceivable for moving the housing body 21. In addition, it is possible to arrange the housing body 21 immovably and to move the medium carrier 1 with respect to the housing body 21. However, moving the housing body 21 with respect to the medium carrier 1 has the advantage that an operator has easier access to the medium carrier 1 in order to feed the sample liquids from the containers mentioned above (not shown) through the through openings 13, 15 into the recess 11 bring.
  • the laser source 17 generates a laser beam 31, for example with a wavelength of 635 nm, which is directed at a predetermined angle, which is preferably of the order of 8 °, onto the interface between the upper part 3 and the medium surfaces 9 facing the lower part 7.
  • the laser beam 31 has a light spot, the area of which corresponds to the individual medium areas 9 and is, for example, 1 ⁇ 5 mm.
  • the laser beam 31 is preferably directed through a prism 33 onto the interfaces mentioned, so that total reflection occurs on an illuminated medium surface 9 and the generated fluorescent light of this medium surface 9 is focused via the imaging optics 19 to the receiver 39.
  • the emitted fluorescent light has a wavelength of 670 nm.
  • the receiver 29 is preferably a photomultiplier.
  • the electrical output signal generated by the receiver 29 is transmitted via a line 35 to an evaluation electronics 37.
  • An aperture 39 is connected upstream of the receiver 29.
  • the laser light reflected by the boundary surfaces m in the form of a beam 41 is preferably absorbed by an absorber 43, so that misinformation which can be attributed to scattered light is avoided.
  • the laser beam 31 is successively focused on the individual medium surfaces 9 by moving the housing body 21 m in the direction of the arrow 23 through the sputtering device 25, 27, with each medium surface 9 advantageously with the same Intensity and uniformity is irradiated.
  • the fluorescent light 45 respectively emitted by the medium surface 9 that has just been irradiated is projected through the imaging optics 19 onto the receiver 35, which sends a corresponding electrical signal to the evaluation electronics 37.
  • the present device makes it possible to use imaging optics 19 one after the other, which compared to the prior art, in which all medium surfaces are illuminated simultaneously and the fluorescent light of all medium surfaces is imaged on a specially designed receiver, can be small.
  • a large number of medium surfaces 90 m in the form of circular dots or pixels on a measurement diumtrager 10 are arranged.
  • the medium surfaces 90 are preferably arranged at the locations of vertically intersecting rows and columns of medium surfaces.
  • To illuminate the medium 90 faces a a light beam 310 is generating light source 170 provided as it is at the preference ⁇ a laser source.
  • the fluorescent light 450 reflected by a medium surface 90 is detected by imaging optics 190 and directed to a receiver 290.
  • the medium transformer 10 is for example similar to the Medi ⁇ umtrager 1 constructed and comprises (not shown) m a recess up to about 10,000 rasterformig arranged point-shaped medium surfaces 90. This medium surfaces 90 can be sequentially rinsed with several liquids at the same time as a whole.
  • the laser source 310 generates a laser beam 310 and focuses it on a medium surface 90.
  • the laser beam 310 has a circular cross section of approximately 50 to 100 ⁇ m corresponding to the surface of the punctiform medium carrier 90.
  • the fluorescent light 450 reflected by excitation from the medium is emitted by the imaging optics 190 onto the sensor of the receiver 290.
  • the medium surfaces 90 arranged in a grid pattern can be illuminated successively and to emit a fluorescent Rescent light 450 can be excited.
  • the light source 170, the imaging optics 190 and the receiver 290 together m arranged in a housing, not shown, which is movable by the drive, which can have, for example, the spindle drive already explained above for moving the housing in the direction of the rows of medium surfaces 90 (arrows 300) and a further spindle drive of the type described above , through which the housing can be moved in the direction of the columns of the medium surfaces 90 (arrows 301).
  • the drive which can have, for example, the spindle drive already explained above for moving the housing in the direction of the rows of medium surfaces 90 (arrows 300) and a further spindle drive of the type described above , through which the housing can be moved in the direction of the columns of the medium surfaces 90 (arrows 301).
  • the advantage of the described embodiment is that two-dimensionally arranged medium surfaces 90 can also be read out without the excitation power being too high or the sensitivity depending on the position. This is due to the point-like scanning of the individual medium surfaces 90.
  • the imaging optics 190 can advantageously be made relatively small and inexpensive, the excitation power only having to be designed for a single point-shaped medium surface 90.

Landscapes

  • Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un équipement d'éclairage et de reproduction, notamment lors de la réalisation de tests d'immunofluorescence quantitatifs. Un faisceau lumineux (31; 310), produit par une source lumineuse (17; 170), est dirigé sur un support d'agent (1; 10) afin de produire une lumière fluorescente (45; 450) sur des plages d'agent (9; 90) du support (1; 10). Une optique de reproduction (19; 190) permet de projeter la lumière fluorescente (45; 450) sur un récepteur optique (29; 290). Le faisceau lumineux (31; 310) est dirigé sur une plage d'agent (9; 90) à la fois et la lumière fluorescente (45; 450) émise par cette plage d'agent (9; 90) est projetée sur le récepteur (29 ; 290) par l'optique de reproduction (19; 190). Un mécanisme permet de produire un mouvement relatif entre le support d'agent (1; 10), d'une part, et la source de lumière (17; 170), l'optique de reproduction (19; 190) et le récepteur (29; 290), d'autre part, aux fins d'éclairage successif des différentes plages d'agent (9; 90).
PCT/DE2001/001301 2000-04-05 2001-04-04 Equipement d'eclairage et de reproduction, notamment lors de la realisation de tests d'immunofluorescence WO2001077645A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DE10017001 2000-04-05
DE10017001.3 2000-04-05
DE10038222 2000-08-04
DE10038222.3 2000-08-04

Publications (1)

Publication Number Publication Date
WO2001077645A1 true WO2001077645A1 (fr) 2001-10-18

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PCT/DE2001/001301 WO2001077645A1 (fr) 2000-04-05 2001-04-04 Equipement d'eclairage et de reproduction, notamment lors de la realisation de tests d'immunofluorescence

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4626684A (en) * 1983-07-13 1986-12-02 Landa Isaac J Rapid and automatic fluorescence immunoassay analyzer for multiple micro-samples
DE19628002C1 (de) * 1996-07-11 1997-12-18 Inst Chemo Biosensorik Vorrichtung und Verfahren zur Durchführung von Fluoreszenzimmunotests
WO1998049543A1 (fr) * 1997-05-01 1998-11-05 Nanogen, Inc. Systeme de detection optique a balayage
US5891656A (en) * 1992-09-14 1999-04-06 Sri International Up-converting reporters for biological and other assays using laser excitation techniques
US6008892A (en) * 1997-05-23 1999-12-28 Molecular Dynamics, Inc. Optical substrate for enhanced detectability of fluorescence

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4626684A (en) * 1983-07-13 1986-12-02 Landa Isaac J Rapid and automatic fluorescence immunoassay analyzer for multiple micro-samples
US5891656A (en) * 1992-09-14 1999-04-06 Sri International Up-converting reporters for biological and other assays using laser excitation techniques
DE19628002C1 (de) * 1996-07-11 1997-12-18 Inst Chemo Biosensorik Vorrichtung und Verfahren zur Durchführung von Fluoreszenzimmunotests
WO1998049543A1 (fr) * 1997-05-01 1998-11-05 Nanogen, Inc. Systeme de detection optique a balayage
US6008892A (en) * 1997-05-23 1999-12-28 Molecular Dynamics, Inc. Optical substrate for enhanced detectability of fluorescence

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