EP1358512A2 - Dispositif d'observation d'echantillons par fluorescence, notamment de facon sequentielle - Google Patents
Dispositif d'observation d'echantillons par fluorescence, notamment de facon sequentielleInfo
- Publication number
- EP1358512A2 EP1358512A2 EP02702453A EP02702453A EP1358512A2 EP 1358512 A2 EP1358512 A2 EP 1358512A2 EP 02702453 A EP02702453 A EP 02702453A EP 02702453 A EP02702453 A EP 02702453A EP 1358512 A2 EP1358512 A2 EP 1358512A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- sample
- objective
- observation
- samples
- support
- 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- Withdrawn
Links
- 238000000799 fluorescence microscopy Methods 0.000 title 1
- 238000012545 processing Methods 0.000 claims description 17
- 238000005497 microtitration Methods 0.000 claims description 14
- 238000006073 displacement reaction Methods 0.000 claims description 8
- 238000007493 shaping process Methods 0.000 claims description 7
- 238000003709 image segmentation Methods 0.000 claims description 3
- 238000012546 transfer Methods 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000000523 sample Substances 0.000 abstract description 23
- 239000012472 biological sample Substances 0.000 abstract description 3
- 230000005855 radiation Effects 0.000 description 23
- 230000003287 optical effect Effects 0.000 description 15
- 230000005284 excitation Effects 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 9
- 238000013519 translation Methods 0.000 description 7
- 230000014616 translation Effects 0.000 description 7
- 238000000018 DNA microarray Methods 0.000 description 4
- 239000003086 colorant Substances 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000011218 segmentation Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 229910052724 xenon Inorganic materials 0.000 description 1
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
- G02B21/04—Objectives involving mirrors
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
Definitions
- the present invention relates to a device for observing samples by fluorescence, in particular sequentially.
- This support is for example a micro-titration plate ("microtiter plate”) at the bottom of the wells ("wells") from which the biological samples are deposited.
- microtiter plate a micro-titration plate
- a particularly important application of the invention relates to high-throughput cellular analysis where cells labeled with a fluorophore are deposited at the bottom of the wells of an icititration plate comprising for example 96 or 384 wells.
- the images of the cells are then formed by observing the wells one after the other. These images are stored and then processed sequentially.
- the object of the invention is in particular to analyze this kind of plate at a high rate, the analysis time of each well being of the order of 1 second to 2 seconds, and thus makes it possible to complete the cellular instrumentation which uses a lot cytometers.
- these are limited to the analysis of cells suspended in a fluid and often only allow a low reading rate.
- biochips are images of biochips (“biochips”) of low complexity from the fluorescence of these biochips, each of these being housed at the bottom of a well of a micro plate. -titration comprising for example 96 wells. It is therefore advantageous to use a large quantity (several tens) of microchips of the MICAM genus (registered trademark), comprising 128 electrodes, and to correlate the results obtained for these chips, for example when studying the expression of the t gene. s.
- the plate supporting the biochips can be provided with various devices, for example means for circulating fluids or means for monitoring (“monitoring") temperatures.
- a device for sequential observation of samples is already known. It is an epi-illumination and fluorescence microscope, equipped with a camera.
- This microscope includes a motorized stage making it possible to move a microtitration plate to successively illuminate the wells and thus successively acquire the images relating to these wells.
- This kind of light source has a wide spectrum but its lifespan does not exceed 200 to
- FIG. 1 schematically illustrates an example of such a known device, intended for the observation of samples 2 placed in the wells
- the samples are excited by radiation 12 from a laser 14 and provide fluorescence radiation 16 as a result of this excitation.
- the bottom of the micro-titration plate is transparent to light from the laser as well as to this fluorescence radiation.
- the light coming from the laser is shaped using an appropriate optic 18 and then filtered using an appropriate filter 20.
- the light thus filtered is reflected, thanks to a dichroic mirror 22, in the direction of the objective 24 of the microscope. It crosses this objective 24 which focuses it on the sample studied. It is specified that the samples are studied one after the other, the micro-titration plate being for this purpose arranged on suitable displacement means, not shown.
- FIG. 1 a camera 26 which is intended to capture the image of the sample studied, thanks to the fluorescence radiation 16 emitted by this sample.
- This fluorescence radiation 16 also passes through the objective 24 of the microscope and then the dichroic mirror 22 (the latter being able to reflect the radiation 12 and transmit the radiation 16) and reaches the camera 26 after having passed through another filter 28 provided for eliminate the light from the laser that may also reach this camera. It should be noted that the use of the dichroic mirror is made necessary, for the separation of the radiations 12 and 16 (whose respective wavelengths are for example equal to 488 nm and to 520 nm), by the existence of a path common to these radiations.
- the filter is not perfect so that a small amount of stray light always reaches the camera.
- Fluorimeters They either use lasers or wide spectrum lamps that are filtered. However, these fluorimeters use a photomultiplier to detect the total emission of fluorescence from a well studied. Consequently, they do not provide any image and therefore cannot be used when one wants to obtain it.
- the subject of the present invention is a device for observing samples by fluorescence, which is quick and simpler than the known device, represented in FIG. 1.
- a device according to the invention is capable of forming 384 fluorescence images in approximately 10 minutes.
- the subject of the present invention is a device for observation, by fluorescence, of at least one sample placed on a support, this device comprising:
- these lighting means comprising at least one light source
- this observation objective being provided for forming an image of the sample when the latter is illuminated
- this device being characterized in that the observation objective is a catadioptric objective and in that the lighting means further comprise means for reflecting the light supplied by the source towards the sample, these reflection means being arranged between the observation objective and the support.
- the catadiotric objective comprises: - a parabolic mirror intended to capture and then reflect light emitted by the sample when the latter receives the light emitted by the source, and - a auxiliary mirror which is placed at the focal point of the parabolic mirror and intended to capture the light reflected by this parabolic mirror and to reflect this light towards the acquisition means, the means for reflecting the light supplied by the source being disposed between the auxiliary mirror and the support .
- the lighting means further comprise means for shaping the light emitted by the source.
- the lighting means comprise a plurality of sources, capable of emitting lights of different wavelengths, and means for activating any of these sources.
- the acquisition means may include a camera with charge transfer device, or CCD camera.
- the device which is the subject of the invention further comprises filtering means arranged between the retro-reflecting objective and the acquisition means and designed to allow only the light emitted by the sample to pass when the latter is illuminated.
- a support capable of receiving a plurality of samples is used and the relative displacement means are provided for successively placing these samples on the observation axis, so as to sequentially observe these samples.
- This support is for example a micro-titration plate comprising a plurality of wells in which the samples are respectively placed.
- the device can also comprise means for automatically positioning the well on the observation axis.
- These positioning means comprise, for example, a four-quadrant photodiode.
- the device which is the subject of the invention may further comprise means for processing each image acquired by the acquisition means.
- the processing preferably includes a step of image segmentation and a step of calculating parameters for each well.
- FIG. 1 is a schematic view of a known device, allowing the observation of samples, and has already been described,
- Figure 2 is a schematic perspective view of a particular embodiment of the device object of the invention
- Figure 3 is a schematic view of different optical means forming part of the device of Figure 2
- Figure 4 schematically and partially illustrates another device according to the invention, using lasers of different wavelengths
- Figure 5 is a schematic and partial view of another device according to the invention, using a four-quadrant photodiode.
- the device that can be seen in these figures is intended for the observation of the samples 2 contained in the wells 4 of the micro-titration plate 6.
- This device comprises an inverted microscope 30 which is equipped for 1 epi-illumination.
- the micro-titration plate 6 comprises for example 384 wells. This plate is held horizontally on a plate support frame 32 which is movable relative to the frame 34 of the inverted microscope.
- a plate 36 is provided which can be displaced in translation relative to the microscope frame 34, in a horizontal direction x, and the plate support frame 32 also forms a plate which is movable in translation relative to the plate 36, according to a another horizontal direction y which is perpendicular to the x direction.
- the device of FIGS. 2 and 3 also includes a microscope optic forming a catadioptric objective 38 also called "reflection objective". This catadioptric objective is carried by the turret (not shown) which comprises the frame 34 of the microscope.
- the device also includes an excitation laser 42 provided for emitting radiation 44 intended to excite the sample 2 that is observed.
- this excitation radiation 44 is shaped by means of an optical shaping assembly 46. It is then reflected, via a mirror 48, towards the well 4 in which is located the sample to be studied 2.
- the excitation radiation reflected by the mirror propagates along the vertical axis Z of the catadioptric objective 38, this axis constituting the optical axis of the microscope. It is specified that, by means of appropriate displacements in the directions x and y, the well, in which the sample to be studied, is placed on this optical axis Z.
- the sample thus excited emits fluorescence radiation 50 which propagates along the Z axis and which is sent, via the catadioptric objective 38, to a camera 52 which the device comprises, after having been filtered by a filter 54 designed to let pass only this fluorescence radiation 50.
- magnification of the catadioptric objective 38 is of the order of 4 to 15 and the camera 52 is a CCD type camera, cooled by appropriate means (not shown).
- FIGS. 2 and 3 show the field-reducing extension ring 56 with which the camera 52 is provided and which is located on the input face of this camera.
- the device of FIGS. 2 and 3 also comprises electronic processing means 58 (computer) intended to process the images supplied by the camera 52 and also to control the latter as well as the displacements in the directions x and y.
- the computer 58 is provided with a video monitor 60 making it possible, in particular, to observe the images acquired thanks to the camera 52.
- a camera comprising 1300 x is used for example 1020 pixels whose size can range from 6.5 mm x 6.5 mm to 10 mm x 10 mm, this camera being cooled to 0 ° C. To form the image of a 3 mm well, this corresponds to a resolution better than 3 ⁇ m. .
- the catadioptric objective used 38 includes a parabolic mirror 62 and a small mirror 64.
- the reflecting face of this mirror 64 is turned towards the mirror 62 and the mirror 64 is placed at the focal point of this mirror 62.
- the mirror 48 intended to reflect the radiation emitted by the laser is a small mirror, which is substantially the same size as the mirror 64 and which is fixed to this mirror 64, above the latter so as to be between the plate 6 and the mirror 64.
- the radiation emitted by the laser 42 in a horizontal direction is reflected by the mirror 48, along the optical axis Z, towards the studied sample.
- This sample is then excited and emits fluorescence radiation 50.
- This fluorescence radiation is picked up by the mirror 62 and reflected towards the mirror 64 which in turn reflects this radiation, along the optical axis Z, towards the camera 52, through the filter 54.
- the parabolic mirror 62 the axis of which is the Z axis, is provided with a central hole 66 which is crossed by this optical axis Z and allows the passage of the fluorescence radiation 50 reflected by the mirror 64.
- FIGS. 2 and 3 An important characteristic of the device in FIGS. 2 and 3 therefore resides in the fact that the light beam emitted by the laser is located outside the optical path in the microscope. This laser beam does not pass through the retro-reflecting objective.
- the beam emitted by the laser is therefore not mixed with the fluorescence radiation. Therefore, a simpler device is obtained than that which is shown in FIG. 1.
- the device of FIGS. 2 and 3 only requires a filter intended to eliminate the stray light capable of being mixed with the fluorescent light.
- the shaping of the beam emitted by the laser, or excitation beam is simpler than in the device of FIG. 1 since it is not focused by the objective of the microscope.
- the optical assembly 46 for shaping the beam 44 emitted by the laser comprises two successive optics 68 and 70, provided to enlarge at least three times the beam emitted by the laser, so as to cover each well studied.
- the beam emitted by the laser has a diameter of the order of 0.8 mm and a divergence of the order of 1 mrad.
- this laser beam 44 enlarged approximately three times by this assembly 46, has a diameter of the order of 2.5 mm.
- a fluorescein (excitation wavelength: 488 ⁇ m - emission wavelength: 520 ⁇ m) and a rhodamine are combined (excitation wavelength: 550 ⁇ m - wavelength of emission: 580 ⁇ m) or the couple cy 3 / cy 5 (cy 3 : 540/570 ⁇ m - cy 5 : 630/670 ⁇ m).
- a device In a device according to the invention, it is easy to mix two or three separate laser beams and to use a multiband filter, which exists in two or three colors, as well as two or three shutters automatically switched by software allowing the successive activation of the lasers emitting the beams, and the acquisition of two or three superimposed images, of distinct colors (or even more if a greater number of lasers are used), and this for each well .
- This is schematically illustrated in FIG. 4 where we see, schematically and partially, a device according to the invention comprising three lasers 72, 74 and 76 designed to emit excitation radiation from samples having lengths d '' different wave (for example 488 ⁇ m, 532 ⁇ m and 550 ⁇ m).
- Each of these lasers 72, 74 or 76 is successively followed by an optical assembly 78, 80 or 82 for shaping the beam supplied by this laser, a shutter 84, 86 or 88 (each shutter being controlled by the computer 58) and a mirror 90, 92 or 94.
- These mirrors are provided to obtain, as a function of the shutter which is actuated, a laser beam directed along an axis Y perpendicular to the optical axis Z and meeting the mirror 48.
- the mirror 94 associated with the laser 76 is a 45 ° mirror, simply provided for reflecting the beam corresponding to this laser along an axis X parallel to the optical axis Z.
- the mirror 92 associated with the laser 74 is arranged above the mirror 94 and designed to transmit the beam reflected by this mirror 94 and to reflect the beam corresponding to the laser 74 along this axis X.
- the mirror 90 associated with the laser 72 is provided for transmitting the beam emitted by this laser along the Y axis and reflecting the light coming from the mirror 92 so that the radiation thus reflected propagates along this Y axis (before being reflected on the mirror 48).
- a buffer memory in the computer 58 associated with the camera 52 for transferring an image of a color there while an image is being acquired in another color.
- the acquisition is carried out in a time between 200 ms and 500 ms.
- the transfer is carried out in less than 500 ms.
- FIG. 5 An example of such a device is schematically illustrated in FIG. 5.
- automatic recognition means 96 comprising a four-quadrant photodiode 98.
- FIG. 5 we see the micro-titration plate 6 which receives the excitation laser beam 44. Part of this beam emerges at the upper end of the well studied 4.
- the laser beam 100 emerging from this upper end has a substantially conical shape.
- the four-quadrant photodiode 98 is arranged above the micro-titration plate 6 so that the optical axis Z of the microscope constitutes the axis of this four-quadrant photodiode 98, the latter thus intercepting the emerging beam 100.
- the four electrical outputs of this four-quadrant photodiode are well balanced if the spot resulting from the emerging beam 100 is circular. However, when the well is well positioned relative to the optical axis Z, this spot is circular. There is therefore a setpoint for immobilizing the translation plates at the desired position: if the laser spot is symmetrical, the intensities of the currents I ⁇ , I 2 , I 3 and I supplied by the diode with four quadrants are equal.
- the sum I of these four currents leaving the photodiode represents the light intensity contained in this laser spot.
- the sum of these four currents can be used either to regulate the laser which emits the beam 44 or as information on the proper functioning of this laser or also for laser safety.
- means can be provided to prevent the protective cover from being removed from the device when the laser is in operation.
- This image processing can be applied in real time or in deferred time. If this treatment is applied in real time, the cell analysis device successively link the following tasks for each of the N wells of the micro-titration plate: (1) displacement on ' well n (l ⁇ n ⁇ N) of the micro-titration plate, (2) image acquisition of this well n, (3) image processing of this well n.
- the device which is the subject of the invention allows cytometric analysis of cells of various types, for example neurons, keratinocytes, fibroblasts and tumor cells.
- the purpose of image processing is to analyze the cells present in the samples and to extract interesting parameters from them.
- image processing can be more or less specific. In all cases, this image processing comprises a step of image segmentation and a step of calculating parameters for each well.
- the segmentation of the image makes it possible to identify and separate the cells from the background.
- the gray level histogram is advantageously used to determine the binarization threshold, and morphology algorithms mathematical.
- the parameters calculated for each well are for example parameters of cell size and shape, the maximum level of fluorescence for each cell and the number of cells.
- connectivity analysis algorithms On this subject, consult the documents [1] and [3] mentioned above.
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Microscoopes, Condenser (AREA)
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0101792A FR2820828B1 (fr) | 2001-02-09 | 2001-02-09 | Dispositif d'observation d'echantillons par fluorescence, notamment de facon sequentielle |
FR0101792 | 2001-02-09 | ||
PCT/FR2002/000435 WO2002065160A2 (fr) | 2001-02-09 | 2002-02-05 | Dispositif d'observation d'echantillons par fluorescence, notamment de facon sequentielle |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1358512A2 true EP1358512A2 (fr) | 2003-11-05 |
Family
ID=8859842
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP02702453A Withdrawn EP1358512A2 (fr) | 2001-02-09 | 2002-02-05 | Dispositif d'observation d'echantillons par fluorescence, notamment de facon sequentielle |
Country Status (5)
Country | Link |
---|---|
US (1) | US20040113095A1 (fr) |
EP (1) | EP1358512A2 (fr) |
JP (1) | JP2004530111A (fr) |
FR (1) | FR2820828B1 (fr) |
WO (1) | WO2002065160A2 (fr) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FI20051329A0 (fi) | 2005-12-27 | 2005-12-27 | Wallac Oy | Laitteisto ja menetelmä näytteiden optista mittausta varten |
US9176062B2 (en) | 2006-12-21 | 2015-11-03 | Koninklijke Philips N.V. | Aperture biosensor with trenches |
US7782454B2 (en) * | 2007-02-13 | 2010-08-24 | Bti Holdings, Inc. | Universal multidetection system for microplates |
US9557217B2 (en) | 2007-02-13 | 2017-01-31 | Bti Holdings, Inc. | Universal multidetection system for microplates |
US8445019B2 (en) | 2007-09-26 | 2013-05-21 | Hamamatsu Photonics K.K. | Microparticle dispersion liquid manufacturing method and microparticle dispersion liquid manufacturing apparatus |
JP5236235B2 (ja) * | 2007-09-26 | 2013-07-17 | 浜松ホトニクス株式会社 | 微粒子分散液製造方法および微粒子分散液製造装置 |
CA3030720A1 (fr) | 2010-08-27 | 2012-03-01 | The Board Of Trustees Of The Leland Stanford Junior University | Dispositif d'imagerie microscopique possedant des proprietes d'imagerie avancees |
EP3850341A4 (fr) * | 2018-09-13 | 2022-06-08 | University Of Massachusetts | Système et procédés d'éclairage par fluorescence libre dichroïque utilisant des lentilles d'objectif réfléchissantes |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE1030582B (de) * | 1953-07-22 | 1958-05-22 | Leitz Ernst Gmbh | Mikroskop fuer Beobachtung mit ultraviolettem Licht |
US4289378A (en) * | 1978-06-21 | 1981-09-15 | Ernst Remy | Apparatus for adjusting the focal point of an operating laser beam focused by an objective |
US4284412A (en) * | 1979-07-13 | 1981-08-18 | Ortho Diagnostics, Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
JPS5633572A (en) * | 1979-08-28 | 1981-04-04 | Fujitsu Ltd | Optical rader device |
JP2749069B2 (ja) * | 1988-04-26 | 1998-05-13 | オリンパス光学工業株式会社 | 蛍光顕微鏡装置 |
SE8900612D0 (sv) * | 1989-02-22 | 1989-02-22 | Jonas Johansson | Vaevnadskarakterisering utnyttjande ett blodfritt fluorescenskriterium |
EP0452963B1 (fr) * | 1990-04-20 | 1996-07-31 | Dainippon Screen Mfg. Co., Ltd. | Système objectif de lentille pour l'utilisation avec un microscope |
WO1998017992A2 (fr) * | 1996-10-25 | 1998-04-30 | Applied Imaging, Inc. | Techniques de formation d'images d'hybridation in situ par fluorescence multicolore (m-fish) utilisant de nombreux filtres multibandes avec superposition d'images |
KR100618502B1 (ko) * | 1998-03-16 | 2006-09-01 | 지이 헬스케어 바이오-사이언시즈 코프. | 공초점 마이크로스코피 영상 시스템에서 사용하기 위한 전자기 방출의 집속 시스템 및 방법 |
US6185030B1 (en) * | 1998-03-20 | 2001-02-06 | James W. Overbeck | Wide field of view and high speed scanning microscopy |
EP1123524A2 (fr) * | 1998-09-30 | 2001-08-16 | Trellis Bioinformatics Inc. | Microscopie a haute capacite |
JP2000151916A (ja) * | 1998-11-12 | 2000-05-30 | Fuji Photo Film Co Ltd | 画像情報読取装置 |
US6130745A (en) * | 1999-01-07 | 2000-10-10 | Biometric Imaging, Inc. | Optical autofocus for use with microtiter plates |
US6654119B1 (en) * | 1999-04-21 | 2003-11-25 | Chromagen, Inc. | Scanning spectrophotometer for high throughput fluroescence detection |
-
2001
- 2001-02-09 FR FR0101792A patent/FR2820828B1/fr not_active Expired - Fee Related
-
2002
- 2002-02-05 JP JP2002564622A patent/JP2004530111A/ja active Pending
- 2002-02-05 US US10/467,777 patent/US20040113095A1/en not_active Abandoned
- 2002-02-05 EP EP02702453A patent/EP1358512A2/fr not_active Withdrawn
- 2002-02-05 WO PCT/FR2002/000435 patent/WO2002065160A2/fr not_active Application Discontinuation
Non-Patent Citations (1)
Title |
---|
See references of WO02065160A3 * |
Also Published As
Publication number | Publication date |
---|---|
FR2820828B1 (fr) | 2003-05-02 |
JP2004530111A (ja) | 2004-09-30 |
WO2002065160A2 (fr) | 2002-08-22 |
WO2002065160A3 (fr) | 2002-12-12 |
US20040113095A1 (en) | 2004-06-17 |
FR2820828A1 (fr) | 2002-08-16 |
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