JPH0566194A - Particle measuring apparatus - Google Patents

Abstract

PURPOSE:To carry out highly accurate measurement by measuring the same sample particles twice. CONSTITUTION:Laser light from laser light sources 2a, 2b illuminates sample particles S at a first A and a second B illuminating position and the scattered light (fluorescence) is detected by light detectors 4a, 4b. The time (to) required for the sample particles S to move from the illuminating positions A to B is estimated in advance through calculation, and when an analogue switch 7 is controlled and turned on after the time (to) elapses from the point at which a signal from the light detector 4a is input, then the light (fluorescence) scattered at the second illuminating position B is detected by the light detector 4b.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a particle measuring device for analyzing sample particles by irradiating individual sample particles in a sample with light and optically measuring scattered light and fluorescence obtained. Is.

[0002]

2. Description of the Related Art A flow cytometer is a device for illuminating a sample liquid flowing at high speed with, for example, a laser beam, and detecting a photoelectric signal due to scattered light or fluorescence to elucidate the characteristic structure of a test particle. Immunology, hematology, oncology,
Used in fields such as genetics.

In the conventional particle analyzer used in this flow cytometer or the like, the test particles such as blood cells passing through the flow passage at the center of the flow cell wrapped in the sheath liquid are irradiated with irradiation light such as laser light. The resulting forward and side scattered light, the shape, size,
It is possible to obtain particle properties such as refractive index. Also,
For the test particles that can be stained with a fluorescent agent, by detecting the fluorescence of the test particles, important information for analyzing the test particles can be obtained.
As disclosed in Japanese Patent No. 318989, there is also proposed an apparatus that increases the measurement parameters by using two bundles of irradiation beams and setting the irradiation positions at two positions.

[0004]

However, in the above-mentioned conventional example, in which the irradiation light is irradiated to two irradiation positions in the distribution direction and the measurement is performed, the first irradiation position and the second irradiation position are different from each other. Since the sample particles to be detected are different, a detection signal for the same test particle cannot be obtained, and particularly when the sample concentration is high, there is a drawback that the accuracy of detection measurement is low.

An object of the present invention is to solve the above-mentioned drawbacks of the conventional example, to accurately detect a large number of measurement parameter signals from the same analyte particle without depending on the sample concentration, and to provide high measurement accuracy. It is to provide a particle measuring device.

[0006]

A particle measuring device according to the present invention for achieving the above-mentioned object is to detect scattered light or fluorescence of irradiation light applied to a specimen particle flowing in a flow cell to detect the specimen particle. A particle measuring device for analysis, comprising: an irradiation unit that irradiates the sample particles with light at a first irradiation position or a second irradiation position that are spaced apart in the flow direction, the first irradiation position, and the second irradiation position. Detecting means for individually detecting scattered light or fluorescence at the irradiation position, and the detecting means only when the sample particles irradiated by the irradiation means at the first irradiation position pass through the second irradiation position. And a control means for controlling the detection means so as to detect scattered light or fluorescence at the second irradiation position.

[0007]

In the particle measuring device having the above-mentioned structure, the scattered light / fluorescence obtained by irradiation of one sample particle when passing through the first irradiation position and the sample particle at the second irradiation position. The scattered light / fluorescence obtained by being irradiated when the light passes through is detected and the information of the sample particle is obtained.

[0008]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described in detail based on the illustrated embodiments. In FIG. 1, laser light sources 2a and 2b that irradiate a laser beam to a first irradiation position A and a second irradiation position B, respectively, facing a flow cell 1 that circulates sample particles wrapped in a sheath liquid in the arrow direction. Are arranged with the optical axes OA and OB perpendicular to the flow direction, and on the optical axis OA and the optical axis OB on the opposite sides of the flow cell 1 respectively, a condenser lens 3a, a photodetector 4a, and a condenser Lens 3b, photodetector 4
b is arranged. The output of the photodetector 4a is connected to the signal processing circuit 5a and the timing control circuit 6, and the outputs of the photodetector 4b and the timing control circuit 6 are connected to the analog switch 7. The analog switch 7 is connected to the signal processing circuit 5b, the output of the signal processing circuit 5a is to the A / D converter 8a, and the output of the signal processing circuit 5b is the A / D converter 8a.
b, the outputs of the A / D converters 8a and 8b are connected to a memory 9, the memory 9 is connected to an analysis operation circuit 10, and the output of the analysis operation circuit 10 is output to an output device 11 such as a display printer. It is connected.

The laser light from the laser light source 2a has an optical axis
When traveling on OA and irradiating the sample particle S at the first irradiation position A, the scattered light or fluorescence is condensed by the condenser lens 3a and received by the photodetector 4a. Photo detector 4a
Is inputted to the signal processing circuit 5a and the timing control circuit 6, the pulse amplitude of the detection signal of the sample particle S is converted into a direct current in the signal processing circuit 5a, and converted into a digital signal in the A / D converter 8a. Then, it is stored in the memory 9. On the other hand, the laser light from the laser light source 2b irradiates the second irradiation position B, and the scattered light or fluorescence from the sample particles is condensed by the condenser lens 3b and received by the photodetector 4b to be converted into an electric signal. And is input to the analog switch 7.

In order to move the sample particles S from the first irradiation position A to the second irradiation position B in advance depending on the interval between the first irradiation position A and the second irradiation position B and the flow velocity of the sheath liquid. The required time to is estimated by calculation. When the timing control circuit 6 controls the analog switch 7 to turn on after time to from the time when the signal from the photodetector 4a is input, the scattered light from the same specimen particle S at the second irradiation position B at that time is controlled.・ Fluorescence is detected by the photodetector 4b,
The detection signal is similarly stored in the memory 9 via the signal processing circuit 5b and the A / D converter 8b.

A large number of sample particles S are obtained by such a method.
For each of the two detection signals, the detection signals are obtained and stored in the memory 9, and the analysis calculation circuit 10 performs the analysis calculation of the sample particle S, and then outputs the result to the output device 11. The analog switch 7 is controlled by the timing control circuit 6 to be kept in the ON state until the sample particle S has completely passed the second irradiation position B, and then returned to the OFF state after the passage.

The laser light sources 2a and 2b may be shared by using a reflection mirror or the like, and, for example, Japanese Patent Laid-Open No.
As disclosed in Japanese Patent No. 318989, the light detectors 4a and 4b may be shared by guiding a light beam.

[0013]

As described above, in the particle measuring apparatus according to the present invention, the same sample particle is detected at the first irradiation position and the second irradiation position, so that the concentration of the sample liquid becomes high. However, it is possible to perform highly accurate measurement.

[Brief description of drawings]

FIG. 1 is a configuration diagram of an embodiment.

[Explanation of symbols]

 1 Flow Cell 2a, 2b Laser Light Source 3a, 3b Condensing Lens 4a, 4b Photo Detector 5a, 5b Signal Processing Circuit 6 Timing Control Circuit 7 Analog Switch 8a, 8b A / D Converter 9 Memory 10 Analysis Operation Circuit 11 Output Device

Claims (1)

[Claims] 1. A particle measuring device for analyzing a sample particle by receiving scattered light or fluorescence of irradiation light applied to a sample particle flowing in a flow cell, the first irradiation being spaced in a flow direction. Irradiation means for irradiating the specimen particles with light at a position or a second irradiation position, detection means for individually detecting scattered light or fluorescence at the first irradiation position and the second irradiation position, and the first The detection means is arranged so that the detection means detects scattered light or fluorescence at the second irradiation position only when the sample particles irradiated by the irradiation means at the irradiation position pass through the second irradiation position. A particle measuring device comprising: a control unit for controlling.