JPH03154850A - Specimen inspecting device - Google Patents

Abstract

PURPOSE:To improve the analytic accuracy by scanning a part to be inspected with plural different-wavelength light beams through a single acoustooptic element, inspecting particles through optical measurement, and increasing parameters which are obtained by performing the measurement once. CONSTITUTION:Liquid to be inspected sheathed with a fast laminar flow of sheath fluid flows through the distribution part 1a of a cell 1. The He-Ne laser beam L1 of a light source 2 is reflected by a mirror 6, passed through a dichroic mirror 4 together with the Ar laser beam L2 of a light source 3, and deflected by the acoustooptic element 5 to irradiate the distribution part 1a. The angle of deflection is varied with a control signal and the beams L1 and L2 are made to scan at right angles to a flow of particles to be inspected. The deflection of the element 5 utilizes the diffraction of light, so the deflection angle depends upon the wavelength of the beams as well. The front scattered light at the distribution part 1a is passed through a stopper 7 and converged by a lens 8, and respective scattered light beams are photodetected by a detector 10 through a BPF 9 to obtain information regarding the element to be inspected from the quantity of photodetection and also obtain information even from the back scattered light. Consequently, new particle analytic data which can not be obtained with single wavelength are obtained and the measurement accuracy is improved.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention, for example in a flow cytometer, irradiates test particles passing through a flow cell with a laser beam or the like and collects optical signals from the test particles. The nature of the particles to be detected,
The present invention relates to a sample testing device that analyzes the structure and the like.

[Conventional technology] A flow cytometer is a cell suspension solution that flows at high speed.
In other words, it is a device that illuminates a sample liquid with a light beam, detects the scattered light and photoelectric signals generated by fluorescence, and elucidates the properties and structure of cells. used in the field. In order to accurately obtain scattered light and fluorescence from microscopic cells, a light beam with high output, low noise, and good focusing properties is required, and a laser beam is generally used.

In a conventional particle analysis device used in this flow cytometer, etc., a particle size of 200 tcm
Irradiation light such as a laser beam is irradiated on test particles such as blood cells wrapped in sheath fluid and passing through a flow section with a small rectangular cross section of 200 um, and the resulting forward and side scattered light is used to It is possible to obtain particle properties such as the shape, size, refractive index, cell diameter, volume, and complexity of the nuclear structure of the sample particles. In addition, for test particles that can be stained with fluorescent agents, important information for analyzing the test particles can be obtained by detecting the fluorescence of the test particles from side scattered light in a direction almost perpendicular to the irradiation light. can be found. For example, it is possible to determine the amount of DNA and RNA by staining, and if a fluorescent dye is reacted with an antigen or antibody, the amount of antigen or antibody can be determined by measuring the fluorescence of the test particle that has caused an antigen-antibody reaction. quantitative measurement becomes possible. Note that the light beam irradiated in these cases needs to have a wavelength that excites the fluorescent dye.

[Problems to be Solved by the Invention] However, in the conventional example described above, the particles to be detected in the fluid are not irradiated with light beams of other wavelengths (they are irradiated with only a light beam of a single wavelength, so the scattered light is Limited to single-wavelength light beams, it is difficult to obtain new particle analysis data and cannot improve accuracy, and the fluorescent dyes that can be used for fluorescence measurements are also limited. It is difficult to perform efficient and accurate analysis of measurements.

SUMMARY OF THE INVENTION An object of the present invention is to eliminate the above-mentioned drawbacks, increase the number of measurement parameters that can be obtained in one measurement, and provide an analyte testing device that allows easy alignment adjustment of the optical system and fluid system.

[Means for Solving the Problems] In order to achieve the above object, the specimen testing device according to the present invention irradiates test particles in a fluid with a light beam,
A sample testing device that analyzes sample particles from optical signals obtained by photometry includes: a flow cell section through which sample particles pass one by one; a plurality of laser light sources each generating a light beam with a different wavelength; The present invention is characterized in that it has a single acousto-optic element for simultaneously making the light beams incident thereon and optically scanning the test area within the flow cell section.

[Operation] The specimen testing apparatus having the above configuration optically scans the test area using a single acousto-optic element with a plurality of lights having different wavelengths, and analyzes the test particles by optical measurement.

[Example] The present invention will be explained in detail based on illustrated embodiments.

Figure 1 shows the configuration and diagram of the optical system for forward scattering and photometry, where l is a flow section 1a in which the particles to be detected and the sheath liquid flow together at high speed.
Toward flow cell 1, which is a flow cell having
A He-Ne laser beam source 2 and an Ar'' laser beam source 3 are provided, and a gicchroic mirror 4 and an acousto-optic element 5 are sequentially placed on the optical path of the Ar'' laser beam L2 emitted from the Ar'' laser beam source 3. The laser beam Ll emitted from the He-Ne laser light source 2 enters the guichroic mirror 4 via the mirror 6, and is reflected by the guichroic mirror 4 in the direction of the acousto-optic element 5. In addition, in order to measure the forward scattered light of the laser beams L1 and L2 caused by the test particles in the flow section la, a stopper 7, a condensing lens 8, a bandpass filter 9a, and a condenser lens 8 are placed across the flow cell 1. 9
A photodetector 10a% 10b is provided. By the way, the bandpass filter 9a and the photodetector 10a, the bandpass filter 9b and the photodetector 10b are paired,
are arranged in parallel. Furthermore, the guichroic mirror 4 has wavelength selectivity to reflect the He-Ne laser beam Ll with a wavelength of around 633 nm and reflect/reflect the Ar'' laser beam L2 with a wavelength around 48 Bnm, and also has a bandpass filter. 9a is He-Ne laser beam L1
The bandpass filter 9b has wavelength selectivity to transmit only the Ar'' laser beam L2.

Figure 2 shows the configuration of the photometric optical system for side scattered light, and this optical system is provided in two stages, upper and lower, for the He-Ne laser beam Ll and the upper 2 Ar laser beams. . On the side of the flow cell 1, a condenser lens 11, a lens 12, an aperture 13, a lens 14, a guichroic mirror 15, 16 having wavelength selectivity, and a mirror 17 are sequentially arranged from the flow cell 1 side. Further, in the reflection direction of the dichroic mirror 15, a wavelength selective filter 18, a lens 19, and a photodetector 20 are provided, and in the reflection direction of the dichroic mirror 16, a wavelength selective filter 21. A lens 22 and a photodetector 23 are provided,
Further, in the reflection direction of the mirror 17, a wavelength selective filter 24, a lens 25, and a photodetector 26 are sequentially arranged. Further, the filter 18 or the filter 21 has wavelength selectivity to selectively transmit only the wavelength light beam reflected by the dichroic mirror 15 or the dichroic mirror 16.

With the sample liquid wrapped in the high-speed laminar flow sheath liquid passing through the flow section 1a in the flow cell 1, a He-Ne laser beam L1 is emitted from the 1e-Ne laser beam light source 2.
．． An Ar″ laser beam L2 is emitted from the Ar″″ laser beam light source 3.He-Ne laser beam L2
is reflected by the mirror 6 and the dichroic mirror 4, is deflected through the acousto-optic element 5, and irradiates the flow section 1a of the flow cell 1, while the Ar'' laser beam L2 is transmitted through the dichroic mirror 4, passes through the acousto-optic element 5, and irradiates the flow section 1a of the flow cell 1. Similarly, it is deflected and irradiates the flow section 1a, and the forward scattered light, side scattered light, and fluorescence from both test particles are measured to analyze the test particles.

The deflection angle of the acousto-optic element 5 is changed by a control signal from a control circuit (not shown), and the He-Ne laser beam Ll is
and Ar'' laser beam L2 are scanned in a direction perpendicular to the flow of the particles to be examined.Since this deflection by the acousto-optic element 5 utilizes light diffraction, the deflection angle is equal to the laser beam incident on the acousto-optic element 5. It also depends on the wavelengths of the beams L1 and L2.Since the wavelength of the He-Ne laser beam Ll is longer than that of the Ar'' laser beam L2, as shown in Fig. 3, the He-Ne laser beam Ll is longer than the Ar'' laser beam L2. It also scans a wide angle range.

The forward scattered light in the flow section 1a is collected by the condensing lens 8 via the stopper 7, and the scattered light of the He-Ne laser beam Ll is transmitted through the bandpass filter 9b and is collected by the photodetector 1.
0. The reflected light of the Ar'' laser beam L2 passes through the band-pass filter 9a and is received by the photodetector 10a, and information regarding the target particles can be obtained from the amount of each received light.A stopper 7 is provided. As a result, the unscattered laser beams L1 and L2 are not directly received by the photodetectors 10a and 10b.

On the other hand, the side scattered light is made into parallel light by the condensing lens 11, and then passes through the lens 12, diaphragm 13, and lens 14, and then is divided into dichroic mirrors 15, 16 and mirrors for each wavelength region depending on the shape of the particles to be examined, etc. 17 and filters 18, 21, 24 and lens 1, respectively.
The light is received by the photodetector 20.23.26 via 9.22.25, and information regarding the target particle can be obtained from the amount of the received light.

In this way, the two types of He-Ne laser beam Ll and Ar'' laser beam L2 with different wavelengths optically scan the target particle, and the scattered light can be photometered and treated as a statistical quantity, so that a single It is possible to obtain new particle analysis data that cannot be obtained when using a single wavelength laser beam, and the measurement accuracy is higher than using only a single wavelength laser beam.

The sample liquid at the time of measurement includes, for example, 3,3°-dipropylthiadicarbocyanine excited by the He-Ne laser beam L1, FITC (fluorescein isothiocyanate) excited by the Ar'' laser beam L2, and PI ( It is also possible to mix fluorescent agents such as Providium iodide.If these fluorescent agents are bound to a specific antibody in advance, if an antigen that reacts with that antibody is present in the sample solution, , it becomes possible to cause an antigen-antibody reaction and label the antigen with a fluorescent agent.

Information regarding a specific antigen can be obtained by irradiating the sample liquid labeled with this fluorescent agent with a He-Ne laser beam Ll and an Ar'' laser beam L2 and measuring the fluorescence intensity.

In measurements using only a laser beam of a single wavelength, the number of fluorescent agents that can be excited by the laser beam of a single wavelength is limited, so it can be measured in one measurement, and the number of antigens, etc., is also limited. By using two types of laser beams with different wavelengths, it is possible to efficiently obtain information regarding a large number of particles to be detected.

[Effects of the Invention 1] As explained above, the specimen testing device according to the present invention scans the test area with a single acousto-optic element using a plurality of light beams with different wavelengths, so that a single type of irradiation light is not required. Multiple parameters that could not be obtained can be obtained in one measurement, and the light irradiation area is widened by optical scanning, so
The tolerance for the alignment of the optical system and the instability of the fluid system is increased, and furthermore, it is advantageous in terms of cost because only a single acousto-optic element is required as the optical scanning means.

[Brief explanation of the drawing]

The drawings show an embodiment of the specimen testing device according to the present invention.
FIG. 2 is a block diagram of a photometric optical system for forward scattered light, FIG. 2 is a block diagram of an optical system for side scattered light/fluorescence photometry, and FIG. 3 is an explanatory diagram of the deflection angle of an acousto-optic element.

Claims (1)

[Claims] 1. In a sample testing device that irradiates a light beam onto test particles in a fluid and analyzes the test particles from optical signals obtained by photometry, there is a flow cell section through which the test particles pass one by one, and the light generated by each. The method is characterized by comprising a plurality of laser light sources having different beam wavelengths and a single acousto-optic element for simultaneously inputting the plurality of light beams and optically scanning the test area in the flow cell section. Sample testing equipment.