JPS62245942A - Particle analyzer - Google Patents

Abstract

PURPOSE:To securely adjust an optical axis by enabling a photodetector to detect the projection image of light with which a flow cell is irradiated. CONSTITUTION:A laser beam L converged on the distribution part 1a of the flow cell 1 through an image forming lens 3 is incident on the photodetector 6 through a condenser lens 4 to obtain information on the size of a particle to be analyzed. Then, 90 deg. scattered light and fluorescent light from the objective particle pass through a condenser lens 7 and are made into parallel light by a condenser lens 10 and the parallel light is reflected by dichroic mirrors 11 and 12 and a reflecting mirror 13 and then incident on photodetectors 16, 18, and 20 through condenser lenses 15, 17, and 19 to obtain information on the shape of the objective particle. When the flow cell 1 is replaced and when a sample flow does not run in the center of the flow cell 1, alignment needs to be performed. Namely, the flow cell 1 is lighted by a lighting optical system composed of a light source 23, a slit 22, etc., while the beam L is cut off and the image of the sample flow is projected on the detector 14 through a lens 7 and a mirror 8, so that the optical axis is adjusted securely by using the projection pattern.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a particle analysis device that can accurately perform 7-alignment between a flow cell and a photometric optical system in a flow cytometer or the like.

[Prior Art] In a conventional particle analysis device used in a flow cytometer or the like, a particle size of 200 JLmX in the center of a flow cell, for example, is
Irradiation light is irradiated onto specimen particles passing through a flow section with a minute cross section of 200 pm while being wrapped in sheath liquid, and the resulting forward and side scattered light is used to determine the shape, size, and refractive index of the specimen particles. It is possible to obtain particle-like properties such as

Adjustment of the optical axis in conventional devices is performed by flowing a sample liquid containing analyte particles together with a sheath liquid while observing the scattered light and fluorescence signals. Therefore, there is a drawback that the alignment state is not clear.

[Object of the Invention] An object of the present invention is to eliminate the drawbacks of the above-mentioned conventional example and to enable reliable optical axis adjustment by visually observing the alignment state or projection pattern displayed on a monitor. The purpose of the present invention is to provide a particle analysis device.

[Summary of the Invention] The gist of the present invention for achieving the above-mentioned object is to irradiate sample particles in a fluid with light from a first direction, and photometrically measure the obtained scattered light and fluorescence through a photometric objective lens. In the apparatus for analyzing sample particles, the sample particles are moved in a second direction perpendicular to the first direction. and an optical system that forms a projected image of the sample particles irradiated from the second direction on a split-type photodetector via a light splitter after passing through the photometric objective lens. This is a particle analysis device featuring:

[Embodiments of the invention] The present invention will be explained in detail based on illustrated embodiments.

FIG. 1 is a block diagram of the optical system. A sample liquid, which has been hydrodynamically focused and is surrounded by a sheath liquid in a high-speed laminar flow, passes through the flow section la of the flow cell 1, and a laser light source 2 is arranged in a direction perpendicular to this flow. There is.

In order to guide the laser beam L irradiated from the laser light source 2 to the circulation part 1a, an imaging lens 3 is arranged on the optical axis O1, and furthermore, an imaging lens 3 is arranged on the optical axis O1 to guide the laser beam L emitted from the laser light source 2 to the circulation part 1a.
111, a stopper 4, a condensing lens 5,
Photodetectors 6 are arranged in sequence.

Further, on the optical axis o2, which is a direction perpendicular to the flow direction of the sample particles and the optical axis 01, from the flow cell l side, a condenser lens 7, a light splitting mirror 8, an aperture 9, a condenser lens 10.
Guicroic mirror with wavelength selective characteristics 11.12
and reflective mirrors 13 are arranged in sequence. At a position conjugate with the aperture 9 on the reflection side of the light splitting mirror 8, there is a so-called CCD.
A split type photodetector 14 such as the like is arranged. In addition, in the reflection direction of the gicroic mirror 10, a condenser lens 15,
A photodetector 16 is provided with a condenser lens 17 in the reflection direction of the gicroic mirror 12. The photodetector 18 further includes a condenser lens 19 in the reflection direction of the reflection mirror 13. Photodetectors 2o are arranged. A condenser lens 21 is located on the optical axis 02 on the opposite side of the condenser lens 7 across the flow cell 1.
, a slit 22, and a light source 23 are arranged.

The laser beam L focused on the flow section la of the flow cell l via the imaging lens 3 is scattered by the sample particles,
The forward scattered light enters the photodetector 6 via the condenser lens 4, and information mainly regarding the size of the sample particles is obtained.

In addition, the 90' scattered light and fluorescence from the sample particles are converted into parallel light by the condenser lens 10 through the condenser lens 7, and the 90' scattered light and fluorescence from the sample particles are converted into parallel light by the condenser lens 10. The light is reflected by the reflecting mirror 13, and enters the photodetector 16, 18, 2o through the condensing lens 15, 17, 19, respectively, and information mainly regarding the shape of the sample particle is obtained. Although not shown in the drawings, barrier filters are often provided in front of each photodetector 16, 18, and 20 to selectively pass light in the desired wavelength range.

In an optical system for extracting side-scattered light and fluorescence, it is desirable that the sample flow and the aperture 9 have an optically conjugate relationship, in order to avoid picking up fluorescence, etc. other than the sample particles. is important.

Therefore, when flow cell 1 is replaced or when the sample flow does not necessarily flow through the center of flow cell 1,
It becomes necessary to perform 7 alignments to satisfy the above-mentioned conjugate relationship.

In order to perform this alignment, the flow cell l is illuminated by an illumination optical system consisting of a light source 23, a slit 22, and a condensing lens 21 while the laser beam L from the laser light source l is cut. The sample stream is projected onto the split photodetector 14 using the split mirror 8 .

FIG. 2 shows the relative relationship between the flow cell I and the split-type photodetector 14, on which the inner wall 1b of the flow section 1a and the sample flow S are projected. Now, if the sample flow S has the property of absorbing the light from the light source 23, then the third
An output signal waveform having a distribution as shown in the figure is obtained, and an absorption band B of the sample flow S appears in the center thereof. If the bit position mN of the split photodetector 14 and the aperture 9 are made to match optically in advance, 70-cell 1 and side scattering φ
By relatively moving the fluorescence optical system, bit position 1α
Alignment can be performed by aligning so that the signal waveform as shown in FIG. 3 takes an extreme value at position N.

In this case, if the wavelength range of the light source 23 is infrared light, the light splitting mirror 8 is a gicchroic mirror that reflects this wavelength range light toward the splitting type photodetector 14 and passes light outside the wavelength range. , photometry and observation can be performed efficiently at the same time without cutting the laser beam L. [Effects of the Invention] As explained above, the particle analysis device according to the present invention can generate a projected image by light irradiated onto the flow cell. The alignment status can be easily observed by detecting it with a photodetector.
Alignment] It! 't'li can be implemented.

[Brief explanation of drawings]

The drawings show an embodiment of the particle analysis device according to the present invention, and FIG. 1 is an optical configuration diagram, FIG. 2 is a relationship diagram between a blow cell and a split-type photodetector, and FIG. 3 is a split-type photodetector. FIG. 3 is a waveform diagram of an output signal from a detector. Symbol l is a flow cell, 1a is a flow section, 2 is a laser light source, 3 is an imaging lens, 5.7.10.21 is a condensing lens,
6 is a photodetector, 8 is a light splitting mirror, 9 is an aperture, 11.1
2 is a guichroic mirror, 13 is a reflecting mirror, 14 is a split type photodetector, 1B, 18.20 is a photodetector, 22 is a slit, and 23 is a light source.

Claims (1)

[Claims] 1. In an apparatus for analyzing sample particles by irradiating light onto sample particles in a fluid from a first direction and measuring the resulting scattered light and fluorescence through a photometric objective lens. an optical system that irradiates the sample particles from a second direction orthogonal to the first direction; and a light splitter after the projected image of the sample particles irradiated from the second direction passes through the photometric objective lens. 1. A particle analysis device comprising: an optical system formed into a split type photodetector via a split type photodetector. 2. The particle analysis device according to claim 1, wherein the light splitter guides light outside the photometric wavelength range to the split type photodetector. 3. Regarding the photometric objective lens, a diaphragm is arranged at a position conjugate with the sample particles, and the diaphragm and the split-type photodetector are arranged at optically equivalent positions via the light splitter. A particle analysis device according to claim 1.