JPH0643090A - Particle analyzer - Google Patents

Abstract

PURPOSE:To obtain the spectrum of an optical signal and to obtain the more detialed information of particles by using a prism and a diffraction grating in an apparatus, which emits light on sample liquid flow containing the particle components such as blood and urine, detects, the signal from the particles and analyzes the particles. CONSTITUTION:The fluorescence from particles undergoes spectroscopy with a prism 28 and a diffraction grating. The light is separated in accordance with wavelengths. The intensity of the obtained fluorescence spectrum is amplified with an image intensifier 30. The intensity in every wavelength is measured with an image sensor 34.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method in which a sample liquid containing particle components such as blood and urine is made to flow in a sheath flow, and the sample liquid flow is irradiated with light to detect a signal from the particle. The present invention relates to a particle analyzer for analyzing a particle, more specifically, to a particle analyzer capable of obtaining a spectrum of an optical signal by using a spectral means such as a prism or a diffraction grating to obtain more detailed particle information. The sheath flow is a flow in which the periphery of a suspension of particles is covered with a laminar sheath liquid so that the particles can be accurately aligned in a line in the center of the liquid flow and passed therethrough. . And as the sheath liquid,
Usually, a diluent or the like is used.

[0002]

2. Description of the Related Art There is an apparatus for illuminating a sample liquid flow containing particles such as stained cells with fluorescence excitation light and detecting fluorescence emitted from the particles to classify and count the particles. A flow cytometer is an example. Further, there is also an apparatus (imaging flow cytometer) capable of capturing a particle image. When measuring the fluorescence emitted from cells in such an apparatus, a wavelength selecting means such as an optical filter or a dichroic mirror is required in order to separate the desired fluorescence from other light. Further, if a plurality of fluorescences having different wavelengths are to be measured, a plurality of photodetectors are required corresponding to them.

Further, Japanese Patent Laid-Open No. 24535/1990 discloses that
Fluorescence from the sample is separated into continuous wavelength components by a spectroscopic means, each wavelength component that has been separated is detected by a one-dimensional photoelectric detector, and the fluorescence intensity distribution with respect to the wavelength of the test particles can be calculated. A meter is disclosed.

[0004]

However, although it is possible to separate light having a large wavelength difference with an optical filter, it is difficult to separate light having a close wavelength. Moreover, the wavelength distribution of light cannot be measured. That is, it is not possible to know which part of the cell emits fluorescence of what wavelength. Of course, it would be possible if the video image was taken by a video camera and the image was analyzed, but the image would have to be taken and processed for each cell, and the device would be complicated.

Further, in the particle analysis device described in Japanese Patent Laid-Open No. 24-35535, since the dispersed fluorescence is weak,
It is difficult for the detector to detect the fluorescence as it is. Although it is possible to increase the fluorescence intensity by increasing the irradiation intensity of the fluorescence excitation light, this causes a problem of damaging the test particles. Further, when using a charge storage type photoelectric conversion element such as a CCD (charge coupled device), if the stored charge is not reset by any method, the fluorescence of all particles passing through the detection region is integrated. I will end up. Since the intervals between particles are not constant, it is necessary to detect the passage of particles and reset the charge each time. It is an object of the present invention to provide a particle analyzer capable of accurately measuring a fluorescence spectrum for each particle even with weak fluorescence.

[0006]

In order to achieve the above object, in the present invention, fluorescence from particles is separated for each wavelength by a spectroscopic means such as a prism or a diffraction grating, and the fluorescence obtained by the spectroscopic means is obtained. It is characterized in that the intensity of the spectrum is amplified by an amplifying means, for example, an image intensifier, and the intensity is measured by an image sensor for each wavelength. As shown in FIGS. 1 and 3, the particle analyzer of the present invention wraps a sample liquid containing particles in a sheath liquid and causes the sheath liquid to flow into the flow cell 16 to form a sheath flow, and the sample liquid flow 18 is irradiated with light. In the device for detecting particles,
Light source 10 for irradiating sample liquid stream 18 with fluorescence excitation light
And a spectroscopic means 28 that obtains a fluorescence spectrum by dispersing the fluorescence emitted in a predetermined direction among the fluorescence emitted from the particles,
An amplifying means 30 for amplifying the fluorescence spectrum obtained by the spectroscopic means 28, an image sensor 34 for detecting each element of the amplified fluorescence spectrum, and a signal of the image sensor 34 is read out and reset each time a particle passes through. And a signal processing means 38.

In the above-mentioned particle analyzer, as shown in FIGS. 4 and 5, there may be a case in which a light detecting means 36 for detecting scattered light emitted from the particle or transmitted light transmitted through the particle is further provided. Further, as shown in FIGS. 6, 8 and 9, a second light source 40 for further irradiating the particles with white pulsed light, and an image pickup means 52 for picking up a white transmitted light image transmitted through the particles. And may be provided.

Further, as shown in FIG.
Is configured to detect forward scattered light and forward fluorescent light, or as shown in FIG. 4, the light detection means 36 is configured to detect forward scattered light and backward fluorescent light, or as shown in FIG. In addition, the light detection means 36 may be configured to detect side scattered light and backward fluorescence. Further, as shown in FIG. 6, the fluorescence spectrum detection system and the particle image capturing system are arranged on the same optical axis, or as shown in FIG. 9, the fluorescence spectrum detection system and the particle image capturing system are orthogonal to each other. In some cases, the irradiation system of the first light source and the irradiation system of the second light source are arranged so as to be orthogonal to each other, as shown in FIG.

Further, another particle analyzer of the present invention is shown in FIG.
As shown in 0, in a device for wrapping a sample liquid containing particles in a sheath liquid and flowing it into the flow cell 16 to form a sheath flow, and irradiating the sample liquid flow with light to detect particles,
The sample liquid is a flat flow 64 that is wide in one direction and narrow in the other direction.
That is, the light source 10 for irradiating the sample liquid flat flow 64 with the fluorescence excitation light and the fluorescence emitted from the wider one of the sample liquid flat flow among the fluorescence emitted from the particles are obtained to obtain the fluorescence spectrum. The spectroscopic unit 28, the amplification unit 30 that amplifies the fluorescence spectrum obtained by the spectroscopic unit 28, the two-dimensional image sensor 70 that detects each element of the amplified fluorescence spectrum, and the two-dimensional image sensor each time a particle passes through. And a signal processing unit 72 that reads out the signal of 70 and resets it.

In the apparatus shown in FIG. 10 described above, a light detecting means 36 for detecting scattered light emitted from particles or transmitted light transmitted through the particles, which is emitted toward the wider width of the sample liquid flat flow, is further provided. May be provided. Further, in the device shown in FIG. 10, as shown in FIG. 12, a second light source 40 for irradiating particles with white pulse light is further provided.
And an image pickup means 5 for picking up a white transmitted light image that has passed through the particles.
2 may be provided.

[0011]

The fluorescence emitted from the particles upon irradiation with the fluorescence excitation light is dispersed by the spectroscopic means 28 to obtain a fluorescence spectrum image. This fluorescence spectrum is amplified by amplification means, for example, the image intensifier 30, and the intensity is measured for each wavelength by the image sensor 34 or 70. When a plurality of one-dimensional image sensors are provided as the image sensor, the fluorescence spectrum of each particle can be simultaneously measured for a plurality of particles. On the other hand, the fluorescence excitation light that has passed through the particles and the scattered light that has been scattered by the particles are detected by the light detection means 36, and the signal processing device 3
At 8 the passage of particles is determined. When the passage of particles is completed, the signal of the image sensor is read out and the image sensor is reset.

[0012]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention will be described in detail below with reference to the drawings. However, the dimensions, materials, shapes, relative positions, etc. of the components described in this example are as follows.
Unless otherwise specified, the scope of the present invention is not intended to be limited thereto and is merely an illustrative example. Example 1 FIG. 1 shows the configuration of a particle analyzer in Example 1. The light source 10 is a fluorescence excitation light source and is Ar or He-C.
d or a laser light source such as a semiconductor laser, or a light source such as a continuous emission type Xe lamp. When a light source having a continuous spectrum such as a Xe lamp is used as the light source, an arbitrary excitation wavelength can be selected by using the wavelength selection filter 12. If a laser is used as the light source, the filter 12 is unnecessary. The condensing lens 14 is for condensing the light from the fluorescence excitation light source 10 into the sample liquid flow 18 flowing through the center of the flow cell 16, and the spot size at the time of condensing is 10 × 2.
It is desirable that the thickness is about 00 μm. Flow cell 16
Is for passing cells through the detection region while aligning the cells one by one by the sheath flow measurement method, and is arranged at the focal position of the excitation light flux narrowed down by the condenser lens 14. The flow cell 16 is made of a transparent material such as glass or plastic, and has a gradually narrowed introduction channel, a narrow measurement channel connected to the introduction channel, and a sheath liquid supply port provided in the introduction channel. And an outlet provided downstream of the measurement flow path.

When the test particles pass through the irradiation region of the fluorescence excitation light, scattered light (forward scattered light) and fluorescence (forward fluorescence) are obtained. These lights are collected by the light receiving lens 22. Two
Reference numeral 0 is a light shielding plate for shielding the direct light from the light source 10. The scattered light is reflected by the dichroic mirror 24 and is incident on the light detecting means, for example, the CCD line sensor 36.
The signal from the line sensor 36 is input to the signal processing device 38, and the passage of particles is detected. Further, the size and number of particles that have passed are also detected. When particles are detected by transmitted light, it is necessary to remove the light shielding plate. On the other hand, the fluorescence passes through the dichroic mirror 24, passes through the slit 26, and enters the spectroscopic means 28. FIG. 2 shows a characteristic diagram of the dichroic mirror 24.

The spectroscopic means 28 is for converting the fluorescence emitted from the cells into a spectrum. By using, for example, a polychromator, a prism, a grating, etc., the fluorescence spectrum image as shown in FIG. 3 is amplified. , On the entrance surface of the image intensifier 30, for example. Reference numeral 58 is a particle. The image intensifier 30 is a photoelectron multiplying element, and is for amplifying the fluorescence spectrum image dispersed by the spectroscopic means 28.
The fluorescence spectrum image incident on the incident surface (photoelectric surface) of the image intensifier 30 is amplified and output to the output surface (fluorescent surface) of the image intensifier 30.
Further, the fluorescence spectrum image output to the image intensifier 30 is formed on the light receiving element (image sensor) 34 by the relay lens 32 or the optical fiber.
By using a CCD line sensor or a photodiode array as the light receiving element (image sensor) 34, the fluorescence intensity for each wavelength is measured. For example, 1
When a CCD line sensor with pixels of 13 μm / 256 pixels is used and the wavelength range from 400 to 656 nm is measured, the fluorescence intensity is measured with a resolution of 1 nm per pixel by appropriately setting the focal length of the spectroscopic means 28. can do.

C as the light receiving element (image sensor) 34
When using a CD line sensor, unlike a photodiode array, since it is a charge storage type sensor, it is necessary to reset the stored charge by some method (if it is not reset, the fluorescence intensity from all the passing particles will be Will be accumulated). Therefore, by using the signal from the line sensor 36, which is a photodetector, the accumulated charge is read out and the charge is reset each time the passage of particles is completed. Further, by processing the signal from the line sensor 36, it is determined whether or not the particle is a particle to be measured. If the particle is not a particle to be measured, the fluorescence spectrum signal from the CCD line sensor 34 is sent to the signal processing device 38. By resetting the data before sending it to, it is possible to fetch only the necessary data. The obtained signal can be processed by the signal processing device 38, and the spectral data for each particle that has passed can be acquired.

In the device of the present invention, since the excitation light and the fluorescence in the light receiving element 34 have different wavelengths, the position of the pixel from which each signal is obtained also differs. Therefore, it is not necessary to use a filter for removing the excitation light. In addition, the flow cell 16
Circular or rectangular slits 2 to limit the detection area inside
It is necessary to install 6. Since the size of the slit 26 is determined by the imaging magnification of the light receiving lens 22, for example, the detection area in the flow cell is φ20 μm, the light receiving lens 2
When the imaging magnification of 2 is 10 times, the size of the slit 26 may be φ0.2 mm. As described above, a flow cytometer device that aims to acquire fluorescence of two or more types of wavelengths using one detection system becomes possible.

Embodiment 2 The apparatus of FIG. 1 is intended to detect forward scattered light and forward fluorescence due to light (fluorescence excitation light) from the light source 10,
Various other examples can be implemented. For example, FIG. 4 shows the configuration of the particle analyzer according to the second embodiment. The apparatus of FIG. 4 differs from the apparatus of FIG. 1 in the arrangement of the irradiation system by the light source 10 (irradiation system of fluorescence excitation light) and the scattered light detection system by the light detection means 36, and the apparatus of FIG. And backward fluorescence is a detection target. Since the excitation light from the light source 10 does not directly enter the fluorescence detection system due to the arrangement of the irradiation system and the mirror 24, highly accurate fluorescence measurement becomes possible.

Example 3 FIG. 5 shows the structure of a particle analyzer in Example 3. The apparatus of FIG. 5 is different from the apparatus of FIG. 4 in the arrangement of the scattered light detection system by the light detection means 36, and the apparatus of FIG. 5 targets side scattered light and backward fluorescence. Since the side scattered light is detected, the light shielding plate 20 is unnecessary. Also in this case, the same effect as that of the device shown in FIG. 4 can be obtained. Further, since the side scattered light is detected, a signal reflecting the difference in the internal structure of the particles can be obtained.

Example 4 FIG. 6 shows the structure of a particle analyzer in Example 4. This example uses the signal obtained in Example 1,
1 shows the configuration of an apparatus that captures a white light image of cells that emit fluorescence of a specific wavelength. Fluorescence excitation light source 10 as a light source
In addition, a pulse emission type light source (for example, a Xe flash lamp) having a wide wavelength range in the visible light range is used as the cell image capturing light source 40. The irradiation light from the light source 40 is collimated by the collimator lens 42 and enters the half mirror 46. The half mirror 46 is the excitation light source 1
0 is used to match the irradiation area of the imaging light source 40, and the ratio of transmitted light to reflected light is the fluorescence receiving system.
Although it can be arbitrarily determined according to the amount of light required in the cell imaging system, the transmittance of light from the excitation light source 10 is increased by setting the transmittance to 90% and the reflectance to 10% to increase the fluorescence intensity. Is desirable. Also, half mirror 4
Reference numeral 8 is for transmitting fluorescence obtained from cells and reflecting cell imaging light, and the ratio of the reflected light to the transmitted light can be determined in accordance with the amount of light required in each system, like the half mirror 46. . The electronic shutter 50 causes the image intensifier 3 to operate when the cell imaging light source 40 emits light.
This is to prevent excessive light from entering 0. Instead of using this electronic shutter, an image intensifier having a gate function may be used.

The image pickup means, for example the CCD camera 52, is for picking up a white light image of cells. However, CC
If the imaging area of the D camera and the irradiation area of the excitation light overlap, the excitation light always enters the CCD camera 52, and the brightness of the CCD element is saturated. Therefore, as shown in FIG. It is necessary to shift the irradiation area 56 of the excitation light source 10 and the imaging area 57 of the CCD camera 52 in advance. 5
8 is a particle. Further, if a light source such as a He-Cd laser that emits light having a wavelength outside the visible region or at the edge of the visible region is used as the excitation light source 10, it does not affect the color imaging of the cell image. The signal processing device 54 processes the signal from the light receiving element (image sensor) 34, determines whether the cell passing through the imaging region is a cell to be measured, and determines that the cell is a target cell. In this case, a trigger pulse for causing the white light image capturing light source 40 to emit light is generated, and the obtained signal is analyzed.

The measurement procedure will be described below. The fluorescence excitation light source 10 constantly irradiates the particle passage area of the flow cell 16 and monitors the passage of cells. When the cells dyed with the fluorescent dye pass through, the fluorescence emitted from the cells and the transmitted excitation light are collected by the light receiving lens 22, pass through the half mirror 48, and the excitation light component is removed by the dichroic mirror 24. , Passes through the circular slit 26 and enters the spectroscopic means 28. The fluorescence that has entered the spectroscopic means 28 is divided into spectra, and then passes through the electronic shutter 50 to form a spectrum image as shown in FIG. 3 on the image intensifier 30. This spectral image is amplified by the image intensifier 30 and output to the phosphor screen of the image intensifier 30. The spectral image output to the phosphor screen of the image intensifier 30 is
An image is formed on the light receiving element 34 by the relay lens 32. On this occasion,
It is also possible to form an image on the light receiving element 34 by using an optical fiber instead of the relay lens 32. Even if the electronic shutter 50 is arranged in the latter stage of the spectroscopic means 28, the same effect can be obtained. Further, even if the electronic shutter 50 is not used, the same effect can be obtained by using the image intensifier with the gate function.

Thereafter, the detected signal is processed by the signal processing device 5
Analyze in 4. Particles are FITC (fluorescei)
In the case of double-staining with nicotinate and phycoerythrin, the particles to be measured are FITC or Phycoer.
The fluorescent wavelengths emitted by the particles are either 530 nm or 570 nm or both because they are stained with ythrin or both. Therefore, the white light image capturing light source 40 is caused to emit light when one of the fluorescence intensities of 530 nm and 570 nm is greater than or equal to a certain value, or when both are greater than or equal to a certain value. Furthermore, the captured particle image is analyzed for each fluorescence wavelength (530 nm, 57
0nm, 3 types of both) are classified and stored in memory. Or
Compared with the preset fluorescence wavelength pattern,
When the wavelength patterns match, the white light image capturing light source 40 is caused to emit light.

In order to capture a still image of a cell, the light emitting time of the white light image capturing light source 40 must be sufficiently short to obtain a still image of the cell. This light emission time is determined by the speed at which cells pass through the imaging region. For example, if the cell passage speed is 1 m 2 / sec, the light emission time is 1
Must be less than μsec. At this time, the electronic shutter 50 is simultaneously operated to prevent strobe light from entering the image intensifier 30. The light emitted from the white light image capturing light source 40 is reflected by the half mirror 46 and is applied to the cells in the flow cell 16. As a result, the light transmitted through the cells is collected by the light receiving lens 22, reflected by the half mirror 48, and imaged on the CCD camera 52. As described above, a white light image of cells that emit fluorescence of a specific wavelength is acquired.

Example 5 The apparatus of FIG. 6 comprises a system for irradiating fluorescence excitation light from a light source 10,
The irradiation system of the pulsed light for particle imaging by the light source 40 is arranged on the same optical axis, and the detection systems for scattered light, fluorescence, and particle transmitted light images are also arranged on the same optical axis, and forward scattered light, forward Although the fluorescence and transmitted light images are to be detected, various other examples are possible. For example, FIG. 8 shows the configuration of the particle analyzer in the fifth embodiment. The device of FIG.
This device is different from the above device in the arrangement of the irradiation system of the fluorescence excitation light by the light source 10, and the side scattered light by the light source 10, the side fluorescent light by the light source 10 and the transmitted light image by the light source 40 are detected. The light-shielding plate 2 for detecting the side scattered light
0 is unnecessary.

Further, in the configuration of the apparatus of FIG. 6, the light source 1
When the light of 0 and the light of the light source 40 are both visible light, since a half mirror or a dichroic mirror that reflects the light from the light source 10 must be used as the mirror 46, the light from each light source can be efficiently used. Well flow cell 16
Can not lead to. However, in the configuration of the apparatus of FIG. 8, since the mirror 46 is not used, the light from the light source 10 and the light from the light source 40 are efficiently supplied to the flow cell 16 respectively.
There is an advantage that it can be irradiated.

Example 6 FIG. 9 shows the structure of a particle analyzer in Example 6. The apparatus of FIG. 9 is different from the apparatus of FIG. 6 in the arrangement of the irradiation system of the pulsed light for particle imaging by the light source 40 and the imaging system of the particle transmitted light image, and the apparatus of FIG. The forward fluorescent light from the light source 10 and the transmitted light image from the light source 40 are detected. The present embodiment is the same as the particle analyzer of the fourth embodiment except that the white light image capturing system is arranged at a position orthogonal to the fluorescence detecting optical system. In this configuration, the half mirror 46 in FIG.
Since it is not necessary to use 48, there is an advantage that the light amounts of the light sources 10 and 40 can be used efficiently. 1
Reference numeral 5 is a condenser lens, 23 is a light receiving lens, and 60 is a signal processing device.

Example 7 FIG. 10 shows the structure of a particle analyzer according to Example 7. The basic configuration of this embodiment is the same as that of the first embodiment. The feature of this embodiment is that the sample liquid flow is a flat flow 64 instead of a round flow, the light receiving element for detecting the fluorescence spectrum image is a two-dimensional image sensor 70 instead of a one-dimensional image sensor, and a slit is used. The point is that the slit 68 has a wide rectangular shape horizontally instead of a circle. FIG. 11 shows
It is a principal part enlarged view of FIG. Since the sample liquid flow 64 is a flat flow, it is possible to increase the number of particle analysis. Further, since the two-dimensional image sensor 70 is used, the spectrum distribution chart for each point in the X direction can be obtained. In addition, in order to obtain a flat sample liquid flow 64 in the flow cell 16, the introduction flow channel of the flow cell 16 has a shape in which only the width in one direction of the flow channel is gradually narrowed.

For example, the measurement area 2 in the flow cell 16
0 × 150 μm, the imaging magnification of the light-receiving lens 22 is × 40
When the number is doubled, the slit 68 in front of the spectroscopic means 28 is 6 ×
0.8 mm, light receiving element (two-dimensional image sensor) 70
If a CCD area sensor with a size of 1 pixel of 40 μm and 150 × 250 elements (150 pixels in X direction, 250 pixels in Y direction) is used, the fluorescence spectrum from cells can be measured over the entire measurement area, and the wavelength resolution is Also CC
The wavelength resolution per D pixel can be 1 nm. Here, the signal obtained from the light receiving element 70 is converted into the signal processing device 7
By treating with 2, the fluorescence wavelength of fluorescence emitted from many cells can be measured at the same time. When the wavelength of fluorescence emitted from cells is limited to a specific wavelength range, for example, FITC (fluorescein) is used.
When isothiocyanate), phycoerythrin, or propidium iodide is used as a fluorescent dye, 530 nm, 570 nm, 610
If a line-type CCD sensor or photodiode array is installed at the Y-axis position corresponding to the wavelength of nm, it is possible to measure only the spectral component of interest. 62
Is a condenser lens, and 66 is a light shielding plate.

Example 8 FIG. 12 shows the structure of a particle analyzer according to Example 8. This example is the same as the device of Example 7 shown in FIG.
A system for capturing a white light image is added. In this configuration example, the signal obtained from the detector (two-dimensional image sensor) 70 is analyzed by the signal processing device 74, and a preset condition (for example, FITC (fluorescein) is used.
cells that are double-stained with isothiocyanate and phycoerythrin, or if either of the fluorescence intensities of 530 nm and 570 nm is above a certain value or both are above a certain value) When the cells emitting light pass, the light source 40 for capturing a white light image is caused to emit light and the CCD camera 5
The cell image is acquired in 2. Reference numeral 44 is a wavelength selection filter. In the case of the embodiment in which the sample liquid is a flat flow, the arrangement of the optical system can be changed in various ways.

[0030]

Since the present invention is constructed as described above, it has the following effects. (1) Fluorescence from particles is separated for each wavelength by a spectroscopic means such as a prism or a diffraction grating, further amplified by an image intensifier, and its intensity is measured by an image sensor for each wavelength. On the other hand, a plurality of fluorescence intensities can be simultaneously measured with high accuracy. Also,
A fluorescence spectrum image can be obtained. (2) Since the light is separated not by the wavelength selection filter but by the spectroscopic means, the light can be separated well even if the wavelengths are close to each other. (3) When the sample liquid flow is a flat flow and a two-dimensional image sensor is used as the image sensor, the fluorescence spectrum of each particle can be measured simultaneously for a plurality of particles.

[Brief description of drawings]

FIG. 1 is a configuration diagram showing an embodiment of a particle analyzer of the present invention.

FIG. 2 is a characteristic diagram of the dichroic mirror in FIG.

3 is a perspective explanatory view for explaining details around a spectroscopic unit in FIG. 1. FIG.

FIG. 4 is a configuration diagram showing another embodiment of the device of the present invention.

FIG. 5 is a configuration diagram showing another embodiment of the device of the present invention.

FIG. 6 is a configuration diagram showing another embodiment of the device of the present invention.

7 is an explanatory diagram showing an irradiation area of an excitation light source and an imaging area of a CCD camera in the flow cell section shown in FIG.

FIG. 8 is a configuration diagram showing another embodiment of the device of the present invention.

FIG. 9 is a configuration diagram showing another embodiment of the device of the present invention.

FIG. 10 is a configuration diagram showing another embodiment of the device of the present invention.

11 is a perspective explanatory view for explaining details around the spectroscopic unit in FIG. 10. FIG.

FIG. 12 is a configuration diagram showing still another embodiment of the device of the present invention.

[Explanation of symbols]

 10 Excitation Light Source 16 Flow Cell 18 Sample Liquid Flow 20 Light-Shielding Plate 26 Slit 28 Spectral Means 30 Amplifying Means (Image Intensifier) 34 Image Sensor 36 Light Detecting Means 38 Signal Processing Device 40 Imaging Light Source 52 Imaging Means (Video Camera) 56 Irradiation area of excitation light source 57 Imaging area of video camera 64 Sample liquid flat flow 66 Light shield plate 68 Slit 70 Two-dimensional image sensor 72 Signal processing device 74 Signal processing device

Claims (12)

[Claims] 1. An apparatus for detecting particles by wrapping a sample liquid containing particles in a sheath liquid and flowing the same into a flow cell (16) to form a sheath flow, and irradiating the sample liquid flow (18) with light to detect particles. A light source (10) for irradiating the sample liquid flow (18) with fluorescence excitation light, and a spectroscopic means (28) for dispersing the fluorescence emitted in a predetermined direction among the fluorescence emitted from the particles to obtain a fluorescence spectrum, An amplification means (30) for amplifying the fluorescence spectrum obtained by the spectroscopic means (28), an image sensor (34) for detecting each element of the amplified fluorescence spectrum, and an image sensor (34) for each passage of particles. Signal processing means (38) for reading and resetting the signal
And a particle analyzer. 2. The particle analyzer according to claim 1, further comprising a light detection means (36) for detecting scattered light emitted from the particles or transmitted light transmitted through the particles. 3. A second light source (40) for irradiating the particles with white pulsed light, and an image pickup means (5) for picking up a white transmitted light image transmitted through the particles.
2. The particle analyzer according to claim 1 or 2, further comprising: 4. The particle analyzer according to claim 2, wherein the light detection means (36) detects forward scattered light and forward fluorescence. 5. The particle analyzer according to claim 2, wherein the light detecting means (36) detects forward scattered light and backward fluorescence. 6. The particle analyzer according to claim 2, wherein the light detecting means (36) detects side scattered light and backward fluorescence. 7. The particle analyzer according to claim 3, wherein the fluorescence spectrum detection system and the particle image capturing system are arranged on the same optical axis. 8. The particle analyzer according to claim 3, wherein the fluorescence spectrum detection system and the particle image capturing system are arranged orthogonally to each other. 9. The irradiation system of the first light source and the irradiation system of the second light source are arranged orthogonal to each other.
The particle analyzer described. 10. A sample liquid containing particles is wrapped with a sheath liquid and allowed to flow into a flow cell (16) to form a sheath flow,
In an apparatus for irradiating light to the sample liquid flow to detect particles, the sample liquid has a flat flow (6) wide in one direction and narrow in the other direction.
4) is a light source (10) for irradiating the sample liquid flat flow (64) with fluorescence excitation light, and the fluorescence emitted from the wider sample liquid flat flow of the fluorescence emitted from the particles. A spectroscopic means (28) for spectroscopically obtaining a fluorescence spectrum, an amplifying means (30) for amplifying the fluorescence spectrum obtained by the spectroscopic means (28), and a two-dimensional image sensor (for detecting each element of the amplified fluorescence spectrum ( 70) and a signal processing means (72) for reading out a signal from the two-dimensional image sensor (70) and resetting it each time a particle passes through, and a particle analyzer. 11. A light detecting means (36) for detecting scattered light emitted from the particles or transmitted light transmitted through the particles, which is emitted toward the wider width of the flat flow of the sample liquid. The particle analyzer according to claim 10. 12. A second light source (40) for irradiating a particle with white pulsed light, and an image pickup means (5) for picking up a white transmitted light image transmitted through the particle.
2) and are provided, The claim 10 or 11 characterized by the above-mentioned.
The particle analyzer described.