WO2015111349A1 - Dispositif d'analyse d'une image de fluorescence multicolore - Google Patents

Dispositif d'analyse d'une image de fluorescence multicolore Download PDF

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Publication number
WO2015111349A1
WO2015111349A1 PCT/JP2014/084018 JP2014084018W WO2015111349A1 WO 2015111349 A1 WO2015111349 A1 WO 2015111349A1 JP 2014084018 W JP2014084018 W JP 2014084018W WO 2015111349 A1 WO2015111349 A1 WO 2015111349A1
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image
fluorescence
light
sample
color
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PCT/JP2014/084018
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English (en)
Japanese (ja)
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高橋　智
禎昭 杉村
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株式会社 日立ハイテクノロジーズ
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    • 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
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • 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
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • 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
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
    • G01N2021/6441—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks with two or more labels

Definitions

  • the present invention relates to an optical analysis / display device.
  • a biological material such as DNA, RNA, protein, cell, or the like is stained multiple times, and the sample is irradiated with light to detect its fluorescence, and a multicolor image is displayed.
  • the present invention relates to an optical measurement / analysis apparatus that displays or analyzes components.
  • an image is generally detected using a fluorescence microscope. From the multiple-stained sample, fluorescence having wavelength bands corresponding to a plurality of fluorescent reagents is generated. In many cases, these images are usually detected, and a plurality of measured images are combined and displayed in the RGB color image space. For example, a cell fluorescence image stained with the first phosphor is used as an R color (red) intensity image, a cell fluorescence image stained with the second phosphor is used as a G color (green), and the third phosphor. A color image is constructed by making the cell fluorescence image stained with B into B (blue) and synthesizing them.
  • Patent Document 1 discloses an apparatus for acquiring an image by switching filter sets
  • Patent Document 2 discloses, for example, that the fluorescence wavelength band is divided into four, and each of them is different almost simultaneously by four two-dimensional detectors. A method for acquiring four fluorescent images is described.
  • Patent Document 1 When measuring a multiple-stained sample, in Patent Document 1, since it is necessary to switch a plurality of filter sets to acquire an image, the time required for the measurement increases. Specifically, in addition to the exposure time of the two-dimensional camera, the movement (rotation) time when replacing the filter set (excitation filter, fluorescence detection filter, excitation / fluorescence separation dichroic mirror), time required for rest, etc. Machine operations are required, and these require time to repeat several phosphors. Therefore, it becomes difficult to simultaneously acquire a plurality of fluorescent images. Since it cannot be detected at the same timing, it is likely to be affected by changes such as fluorescence fading. It is also difficult to use for objects whose fluorescence changes over time.
  • the fluorescence wavelength band is divided into four, and four different fluorescence images are acquired almost simultaneously with four two-dimensional detectors.
  • four expensive detectors are used, and the apparatus cost is reduced. Becomes larger.
  • the filter set and the like tend to be expensive.
  • measurement is performed by dividing into a plurality of fluorescent wavelength bands, but these wavelength bands basically do not overlap, so in the case of a general phosphor whose fluorescent spectrum is spread over multiple wavelength bands In this case, only a part of the total emission intensity is detected as the detected fluorescence intensity, and detection loss is likely to occur.
  • it is difficult to separate and detect phosphors having close wavelength bands it is generally considered that there are about 3 to 4 phosphor types that can simultaneously acquire a fluorescence image.
  • the present invention provides an apparatus capable of easily and simultaneously measuring a plurality of fluorescent images of a multiple-stained sample.
  • a multi-color fluorescence image analyzer that irradiates a sample stained with a plurality of phosphors with light from a light source and detects and displays fluorescence generated from the sample, A light source for photoexcitation, An irradiation optical system that collects light from the light source and irradiates the sample, A fluorescence condensing system that collects light to detect fluorescence emitted from the sample; A separation unit that divides the collected light at different ratios; An imaging optical system for imaging each of the divided lights; A two-dimensional detector having a plurality of detection pixels for detecting each of the divided and formed images, an addition image forming processing unit for adding the respective detection images, and a ratio of one image to the added image And a data processing unit including an image formation processing unit that generates a color composite image with the ratio image intensity as a hue and the added image intensity as a brightness.
  • the present invention it is possible to provide an apparatus capable of easily and simultaneously measuring a plurality of fluorescent images of a multiple-stained sample.
  • FIG. 3 is a diagram illustrating the principle of simultaneous multicolor measurement in the first embodiment.
  • FIG. 6 is a characteristic diagram of a multi-color simultaneous detection ratio split mirror used in the first embodiment.
  • the block diagram of the measuring apparatus of Example 1 is shown in FIG.
  • the illustrated apparatus has a configuration similar to a microscope, and irradiates a fluorescently-labeled sample 2 with excitation light, detects the generated fluorescence with a two-dimensional detector, and displays data / images.
  • Normal microscope functions such as transmitted illumination, phase contrast image measurement, differential interference image measurement, ND filter, objective lens revolver, XY stage, focusing Z adjustment mechanism, eyepiece, microscope housing, various power supplies, automatic The adjustment mechanism and the like are omitted from the drawing. The same applies to the temperature control vessel heater panel used as necessary.
  • the light from the light source 10 for exciting the fluorescence is condensed by the lens 11, and the light in the necessary wavelength band is taken out by the excitation / fluorescence separation dichroic mirror unit 13 (including the excitation filter, dichroic mirror, and fluorescence filter), The light is reflected and condensed through the objective lens 12 onto the sample 2 held on the sample stage 1 and irradiated.
  • the light source 10 for exciting the fluorescence an ultra-high pressure mercury lamp, a xenon lamp, various laser devices, a high-intensity LED light source, and the like can be used. It can also be introduced via a light light guide.
  • Fluorescence emitted from the sample is collected again by the objective lens 12, the excitation light component such as scattered light is reflected by the dichroic mirror unit 13, and the fluorescence component is taken out by the fluorescence filter.
  • a normal dichroic mirror unit 13 is used. More preferably, the dichroic mirror for excitation / fluorescence separation can be excited in a plurality of wavelength bands and can detect fluorescence in a plurality of wavelength bands. Mirror units are preferred, and dual band, triple band, and quad band filter sets are used.
  • the transmitted fluorescence is transmitted through the auxiliary filter 14 and is incident on the ratio split mirror 15.
  • the two-part mirror for example, has a transmittance characteristic that the transmittance at 450 nm or less is almost 0, 700 nm or more is 100%, and there is a transmittance characteristic that changes almost linearly between them, and it is gentle over a wide wavelength range such as 50 nm to 300 nm.
  • a mirror having transmission characteristics Unlike ordinary dichroic mirrors, aiming for fluorescence from a specific phosphor, the specific wavelength band is not 100% transmitted and the others are not reflected (0% transmitted), but the fluorescence is divided into two parts. Dividing by changing the ratio.
  • fluorescence having a central wavelength of 500 nm is divided and formed into a transmitted fluorescent image having a transmittance of 20% and a reflected fluorescent image having a reflectance of 80%, and fluorescence having a central wavelength of 600 nm is transmitted with a transmittance of 60%.
  • the fluorescence having a central wavelength of 700 nm is divided into a transmitted fluorescent image with a transmittance of 100% and a reflected fluorescent image with a reflectance of 0%.
  • These transmitted fluorescent image and reflected fluorescent image are imaged on two-dimensional detectors 20 and 21 (high-sensitivity cooled two-dimensional CCD camera, cMOS camera, etc.) through imaging filters 18 and 19 through auxiliary filters 16 and 17, respectively. And detect.
  • the control PC (not shown) controls the setting of the exposure time of the two-dimensional detector, the capture timing of the fluorescence image, and the like via a camera controller (not shown).
  • the auxiliary filters 14, 15, and 16 are used as necessary. For example, a short wavelength cut filter is used to further cut out leakage of excitation light that has not been sufficiently cut.
  • the functions of a band pass filter and a long wavelength cut filter are provided.
  • an ND filter for attenuating the light intensity is inserted in order to balance the divided light intensity.
  • a broadband bandpass filter for fluorescence detection may be used.
  • a notch filter for laser removal can be inserted.
  • a cooled high-sensitivity camera or the like as a two-dimensional detector.
  • a cooled cMOS camera having a pixel size of 6.5 ⁇ 6.5 micrometers and a pixel number of 2048 ⁇ 2048 pixels is used.
  • a high sensitivity camera such as a CCD camera can be generally used.
  • CCD camera can be generally used.
  • CCD camera can be generally used.
  • CCD camera can be generally used as back-illuminated and front-illuminated CCD cameras depending on the structure, and both can be used.
  • An electron multiplying CCD camera having a signal multiplying function inside the element is also effective in achieving high sensitivity.
  • the sensor is preferably a cooling type, and by setting the temperature to about ⁇ 20 ° C. or less, dark noise of the sensor can be reduced and measurement accuracy can be improved.
  • the measured transmission fluorescence image and reflection fluorescence image are converted into a color image by the next data processing unit 22.
  • each of the transmitted fluorescent image and the reflected fluorescent image is subjected to a process such as smoothing, and then baseline correction is performed to subtract the dark noise, offset, and background light intensity of the camera.
  • Camera dark images in advance for the processing. A background light image when there is no phosphor is acquired, and processing is performed using them.
  • the exposure time or sensitivity of the two detectors are different, correct them. This is not particularly necessary when the same type of detector is used and detected with the same exposure time.
  • the processed transmission fluorescent image and the reflected fluorescent image are added for each pixel by the addition image formation processing unit.
  • the added image is an image of the sum total of the entire wavelength range of the detected fluorescent light intensity. It is not an image separated for each wavelength, but an intensity image of the entire wavelength range to be measured.
  • a ratio image is obtained from the transmitted fluorescent image and the reflected fluorescent image.
  • the ratio image is (transmission fluorescence image) / (transmission fluorescence image + reflection fluorescence image), that is, (transmission fluorescence image) / (addition image).
  • the ratio image may be (reflected fluorescent image) / (transmitted fluorescent image + reflected fluorescent image), that is, (reflected fluorescent image) / (added image).
  • the ratio image intensity is set as the hue
  • the added image intensity is set as the brightness
  • the saturation is fixed, and is developed in the HSV color space to form a color composite image.
  • the ratio (0.00 to 1.00) may be converted to a hue of 0.0 to 360.0 degrees, or may be converted to 0.0 to 240.0 degrees. If converted to 0.0 to 240.0 degrees, it is in red ⁇ orange ⁇ yellow ⁇ green ⁇ blue ⁇ purple, and the change in wavelength color and the sensation are the same, so normally it is converted at 0.0 to 240.0 degrees. Of course, even if converted from 240.0 to 0.0 degrees, the display color order is only reversed, so there is no problem.
  • FIG. 2 is a diagram for explaining the principle of multicolor simultaneous measurement according to the first embodiment.
  • FIG. 2a is a model image of the sample
  • FIG. 2b is a ratio-divided mirror characteristic
  • FIG. 2c is a detected ratio-divided image
  • FIG. 2d is a ratio image and added image obtained by the data processing unit.
  • FIG. 2e is a color image constructed by the data processing unit.
  • Fig. 2a Consider an image containing six different phosphors as a sample (Fig. 2a). When excited using a light source such as a mercury lamp, six different types of fluorescence are generated, from the left to blue, blue-green, green, yellow-green, orange, and red. These fluorescences are collected and divided by a ratio-divided mirror. As shown in Fig. 2b, the ratio of the split mirror is almost linearly gradual, with transmittance below 450nm being almost 0%, 620nm and above being almost 100%, and between 450 and 620nm from almost 0% to almost 100%. It has the transmittance characteristic to be.
  • the blue fluorescence is reflected with almost no transmission, so that the signal is weak in the transmission image and strong in the reflection image.
  • the fluorescence wavelength is slightly greener than blue
  • the signal is slightly stronger in the transmitted image than in the blue image, and slightly smaller in the reflected image.
  • the transmitted image intensity is slightly smaller than the reflected image intensity
  • the transmitted image intensity is slightly larger than the reflected image intensity. In the case of red, since it almost transmits, the reflected image becomes dark (FIG. 2c).
  • the background light intensity is subtracted, and rotation correction and trimming correction for superposition are performed to obtain an added image. Since the signals other than the six fluorescent light emitting areas are small, there is no point in calculating the ratio at that portion. Therefore, the ratio image for which the signal intensity level of the added image exceeds about 10% is calculated. Is obtained (FIG. 2d).
  • the ratio and hue correspond to each other as shown in FIG. 2f and the intensity of the added image corresponding to the lightness by the image formation processing unit, the multicolor label can be obtained from the transmitted fluorescent image and the reflected fluorescent image detected by the ratio bisection.
  • the sample image can be displayed in color.
  • Table 1 is a characteristic table showing the phosphor types used, their excitation wavelengths, and fluorescence wavelengths.
  • FIG. 3 shows the characteristics of the excitation / fluorescence separation dichroic mirror unit.
  • FIG. 3a shows the excitation light filter characteristics
  • FIG. 3b shows the fluorescence detection filter characteristics
  • FIG. 3c shows the excitation / fluorescence separation dichroic mirror characteristics.
  • the fluorescence wavelength in the sample is distributed with a width of about 300 nm from about 420 nm to about 700 nm.
  • a triple-band dichroic mirror unit was used to excite these simultaneously and detect fluorescence.
  • the ratio bisection mirror shown in FIG. 4 was used.
  • FIG. 5 shows the measured transmission fluorescence image (FIG. 5a) and reflection fluorescence image (FIG. 5b), the ratio image obtained by image processing (FIG. 5c), and the color image (FIG. 5d).
  • the color was set by setting the ratio 0.0 ⁇ 1.0 to the hue 0.0 ⁇ 240 degrees.
  • FIG. 5e shows the ratio distribution of the individual phosphors and shows that the six-color phosphors can be detected separately.
  • the ratio split mirror is not limited to the above. It is sufficient if the transmittance at a certain wavelength in the wavelength range to be measured is substantially different from the transmittance at other wavelengths.
  • the transmittance of the phosphor with the shortest wavelength is 0%, and the transmittance of the phosphor with the longest wavelength is almost 100%. That's fine.
  • the transmittance of the phosphor in the shortest wavelength region is almost 100%, and the transmittance of the phosphor in the longest wavelength region is almost 0%, and the same effect can be obtained.
  • the wavelength range may be further expanded so that the 0% transmission wavelength is shorter and the 100% transmission wavelength is longer.
  • the phosphor species that can be measured simultaneously The number of multiple colors that can be measured simultaneously can be increased.
  • the above 100% does not mean that the transmittance is exactly 100%, but means 100% when the sum of the reflectance and the transmittance is 100%, and the transmittance as an optical substrate. It is not something that restricts.
  • the transmittance as the optical substrate may be 98%, 95%, etc., for example. The same applies to the transmittance of 0%.
  • the transmittance of the phosphor in the shortest wavelength range is approximately 1%
  • the transmittance of the phosphor in the longest wavelength range is approximately 95%. There is an effect.
  • ⁇ Fluorescence in a wide wavelength range of 300 nm can be detected and identified with one type of filter set, and the cost of the optical filter can be reduced. Even if the type of phosphor is changed, the possibility of using the same filter set is hard and versatile.
  • the stability of the device is increased. Moreover, it can measure simply.
  • the apparatus can be configured inexpensively and simply.
  • the fluorescence emitted from the sample is transmitted through the dichroic mirror unit 13 and then detected by switching the total intensity between the transmission side and the reflection side, so that the total intensity can be detected without loss and high sensitivity. It is also effective for conversion.
  • the process was performed on six types of phosphors, but it can be applied to three types, four types, five types, and six or more types of phosphors, and is effective for simultaneous measurement such as multiple staining.
  • the normal fluorescence microphotometry is used.
  • all the detection methods as an image have the same effect.
  • the same effect can be obtained in the image measurement with the confocal irradiation imaging system and the evanescent illumination.
  • the multicolor simultaneous measurement apparatus in the second example shows the results in FIG.
  • Fig. 6a is a transmitted fluorescence image
  • Fig. 6b is a reflected fluorescence image
  • Fig. 6c is a color conversion image. In this way, measurement of multiple stained cell images allows simultaneous measurement of multiple phosphors, enabling simple measurement.
  • Example 2 The same effect as in Example 1 is obtained even with a multiple-stained cell sample.
  • Fig. 7 shows the configuration of another multicolor fluorescence simultaneous measurement apparatus.
  • Example 1 transmitted illumination, phase difference image measurement, differential interference image measurement, ND filter, objective lens revolver, XY stage, focus adjustment Z adjustment mechanism, eyepiece, microscope case, Various power supplies, automatic adjustment mechanisms and the like are omitted from the figure. The same applies to the temperature control vessel heater panel used as necessary.
  • a light source 10 such as an ultra-high pressure mercury lamp, xenon lamp, various laser devices, or a high-intensity LED light source for exciting fluorescence is condensed by a lens 11, and a dichroic mirror unit 13 for excitation / fluorescence separation (filter for excitation, Light including a necessary wavelength band is taken out by a dichroic mirror and a fluorescent filter), reflected, condensed, and irradiated onto the sample 2 held on the sample stage 1 through the objective lens 12.
  • a light source 10 such as an ultra-high pressure mercury lamp, xenon lamp, various laser devices, or a high-intensity LED light source for exciting fluorescence is condensed by a lens 11, and a dichroic mirror unit 13 for excitation / fluorescence separation (filter for excitation, Light including a necessary wavelength band is taken out by a dichroic mirror and a fluorescent filter), reflected, condensed, and irradiated onto the sample 2 held on the sample stage 1 through the objective lens 12.
  • the fluorescence emitted from the sample is collected again by the objective lens 12 to be converted into a parallel light beam, the excitation light component such as scattered light is reflected by the dichroic mirror unit 13, and the necessary fluorescence wavelength component is transmitted by the fluorescence filter. .
  • the transmitted fluorescence is transmitted through the auxiliary filter 14, once imaged by the imaging lens 101, again converted into parallel light by the lens 102, and incident on the ratio split mirror 103.
  • the two-part mirror for example, has a transmittance characteristic of approximately 0% at 420 nm or less, 100% at 600 nm or more, and a transmittance characteristic that changes almost linearly between them, and has a gentle transmission characteristic over the 180 nm wavelength range. It is a mirror. Similar to Example 1, it is possible to simultaneously detect multicolor fluorescence that emits fluorescence in the wavelength range of about 400 nm to 600 nm.
  • the transmitted light from the ratio-divided mirror 103 is reflected by the mirror 104 and the mirror 105 and is imaged by the lens 109 through the auxiliary filter 107.
  • the light reflected by the ratio-divided mirror 103 is reflected by the mirror 106 and imaged by the lens 109 through the auxiliary filter 108.
  • the optical axes of transmitted light and reflected light with respect to the lens 109 are arranged differently as shown in the figure. As a result, the positions of the transmitted light image and the reflected light image when formed by the lens 109 can be shifted, and the two two-dimensional detectors 110 detect both images simultaneously.
  • the measured fluorescent image is converted into a color image by the data processing unit 111.
  • the transmitted fluorescence image and the reflected fluorescence image are separated from the image of the two-dimensional detector. Thereafter, a color image is formed from the transmitted fluorescent image and the reflected fluorescent image divided into two at the same step as in the embodiment, and output and displayed on the display device 112 such as a PC monitor.
  • the measurement field of view becomes smaller than when each image is detected with separate two-dimensional detectors.
  • the number of two-dimensional detectors can be reduced to one, the cost can be reduced.
  • the configuration of the present embodiment has the same effect as described in the first embodiment.
  • Fig. 8 shows another combination in multi-color simultaneous detection.
  • FIG. 8e An example of a filter set when Qdot (R) is used as a fluorescent reagent is shown.
  • the excitation light filter characteristic diagram 8a When eight types are used as Qdot (R) as shown in FIG. 8e, the excitation light filter characteristic diagram 8a, the fluorescence detection filter characteristic diagram 8b, and the excitation / fluorescence separation dichroic mirror characteristic diagram 8c are used.
  • the ratio division filter has a transmittance around 510 nm at the short wavelength end of the transmission wavelength band of the fluorescence filter, based on the emission characteristics of Qdot (R) 525, the characteristics of the fluorescence filter, and the characteristics of the dichroic mirror.
  • the characteristics are 0% and 100% near 705 nm of the fluorescence maximum wavelength of Qdot (R) 705.
  • eight types of Qdot (R) can be simultaneously identified and detected by the same filter set. All can be detected by one exposure, and measurement can be easily performed. Since eight types can be measured simultaneously, an image can be measured at high speed, and high-speed repeated measurement can be performed even in a system that changes over time.
  • the combination of the present embodiment has the same effect as described in the first embodiment.

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Abstract

L'invention concerne un dispositif d'analyse d'image de fluorescence multicolore présentant ce qui suit : une source lumineuse de photoexcitation ; des optiques d'éclairage qui captent la lumière de ladite source lumineuse et éclairent un échantillon ; des optiques de collecte de fluorescence qui collectent de la lumière en vue de détecter la fluorescence ou analogue émis par l'échantillon ; une unité de séparation qui divise la lumière collectée à l'aide de différents rapports ; des optiques d'imagerie qui forment des images respectives à partir de la lumière divisée ; un détecteur à deux dimensions équipé d'une pluralité de pixels de détection pour détecter les images formées respectives ; et une unité de traitement de données qui présente une unité de traitement de formation d'image ajoutée qui ajoute les images détectées les unes aux autres, une unité de traitement de formation d'image de rapport qui calcule le rapport entre une image et l'image ajoutée et une unité de traitement de formation d'image qui génère une image colorée de synthèse à l'aide de l'intensité de l'image de rapport comme composante de nuance correspondante et l'intensité de l'image ajoutée comme composante de valeur correspondante.
PCT/JP2014/084018 2014-01-27 2014-12-24 Dispositif d'analyse d'une image de fluorescence multicolore WO2015111349A1 (fr)

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WO2018150559A1 (fr) * 2017-02-20 2018-08-23 株式会社日立ハイテクノロジーズ Système et procédé d'analyse
US10399964B2 (en) 2017-03-30 2019-09-03 The University Of Kansas Coumarin-linked taxanes for detection and circumvention of cellular efflux
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WO2021070259A1 (fr) * 2019-10-08 2021-04-15 株式会社日立ハイテク Dispositif d'analyse et procédé d'analyse
CN113109314A (zh) * 2021-05-28 2021-07-13 上海睿钰生物科技有限公司 多荧光信号检测系统和方法
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CN113646877A (zh) * 2019-03-28 2021-11-12 浜松光子学株式会社 检查装置及检查方法
CN117402721A (zh) * 2023-11-03 2024-01-16 苏州思迈德生物科技有限公司 一种多色荧光检测的检测装置及检测方法

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