JPS62112035A - Particle analyzing instrument - Google Patents

Abstract

PURPOSE:To make a data analysis in real time with each one piece of specimen particles without requiring intricate devices by providing an interface part to a signal processing system and using a microcomputer. CONSTITUTION:The laser beam L condensed to a flow part 1a of a flow cell 1 via an imaging lens 3 is scattered by the specimen particle and the forward scattered light thereof is made incident on a photodetector 5 via a condenser lens 4. The 90 deg. scattered light and fluorescence from the specimen particle are reflected by dichroic mirrors 7, 8 and a half mirror 9 by each of wavelength region via a condenser lens 6 and is made incident on photodetectors 11, 13, 15. The outputs of the photodetectors are outputted to an analog processing part 20 after amplification and are subjected to calculations such as peak detection, time integration and logarithmic amplification. The calculated signal is subjected to A/D conversion in the interface part 21 and is inputted to a microcomputer part 22. The result of the calculation is displayed on a display device 25.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention is an information processing system that uses a microcomputer to collect optical signal data of light scattered by sample particles generated at short intervals in a flow cytometer or the like. The present invention relates to a particle analysis device that performs high-speed analysis.

[Conventional Branch Technique] In a conventional particle analysis device used in a flow cytometer, etc., for example, 200 ALmX
Stream II with a minute cross section of 200 tiles! Irradiation light is irradiated onto the specimen particles passing through the first part while being wrapped in the sheath liquid,
The resulting forward and side scattered light makes it possible to obtain particulate properties such as shape, size, refractive index, etc. of the analyte.

Since sample particles are passed through the flow section at a rate of 5,000 or more per second to collect and analyze data, conventionally a large computer (minicomputer or larger) is used, but this processing method is Operation is complicated;
Another drawback is that it is expensive.

As one solution to this problem, a device has been developed that incorporates a squared microcomputer into a processing device and performs data analysis. However, these devices temporarily accumulate a large number of data before inputting it to the microcomputer, and then output it all at once to the microcomputer, making real-time simultaneous processing impossible and requiring a large amount of storage space. Therefore, there are problems such as a complicated circuit configuration.

[Objective of the Invention] The object of the present invention is to provide an interface section in the signal processing system and use a microcomputer to perform data analysis for each sample particle in real time without requiring complicated equipment. The object of the present invention is to provide a particle analysis device that can obtain the desired results.

[Summary of the Invention] The gist of the present invention to achieve the above-mentioned object is to provide an irradiation optical system that irradiates a laser beam to sample particles flowing through a flow section in a flow cell, and a method for photometric measurement of scattered light from the sample particles caused by the laser beam. and a signal processing system that analyzes multiple types of signals obtained from the photometric optical system,
The signal processing system performs A/D conversion on the plurality of types of analog signals that are input simultaneously each time one specimen particle passes through, converts them into digital signals, and converts the analog signals into digital signals, which are then processed into data by a microcomputer unit having an arithmetic unit and a storage unit. This is a particle analysis device characterized in that analysis is performed in the analysis FrR.

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

FIG. 1 is a block diagram of an optical system and a signal processing system.

The sample particles that have been hydrodynamically focused pass through the flow section la of the flow cell 1, surrounded by a high-speed laminar flow, and 7g of laser light is emitted in a direction perpendicular to this flow.
2 is placed. In order to guide the laser beam L irradiated from this laser light source 2 to the circulation part 1a, the optical axis 0
An imaging lens 3 is placed on one side, and a condenser lens system 4 is placed on the optical axis 01 on the opposite side of the imaging lens 3 across the flow cell 1 in order to measure the forward scattered light from the sample particles.
and photodetectors 5 are arranged in sequence. In addition, in order to determine M1 the 90" scattered light and fluorescence from the sample particles, a condenser lens 6 is placed from the flow cell 1 side on the optical axis 02, which is a direction perpendicular to the flow direction of the sample particles and the optical axis 01. ,
A guichroic mirror 7.8 having wavelength selection characteristics and a half mirror 9 are arranged in sequence. In the direction of reflection of the microink mirror 7, there is a condenser lens lO1 and a photodetector 1.
1 is a condenser lens 12 and a photodetector 13 in the direction of reflection of the microink mirror 8, and a condenser lens 14 in the direction of reflection of the half mirror 9. Photodetectors 15 are arranged.

As a signal processing system, the outputs of the photodetectors 5, 11, 13, and 15 are connected to amplifiers 16 to 19, respectively, and the outputs of each of these amplifiers 16 to 19 are connected to the analog processing section 2.
Connected to 0 and 5. The output of this analog processing section 20 is the first
interface part? It is connected to the microcomputer section 22 via (2). The microcomputer section 22 is composed of a second interface section 23, a microcomputer main body 244, and a display section 25 from the input side. Furthermore, the output of the analog processing section 20 is sent to the monitor section 26.
It is connected to a single particle counting section 27.

The laser beam L focused on the flow section 1a of the flow cell 1 via the imaging lens 3 is scattered by the sample particles,
The forward scattered light enters the photodetector 5 via the condensing lens 4, and information mainly regarding the size of the sample particles is obtained.

Further, the 90' scattered light and fluorescence from the sample particles pass through the condenser lens 6 and are reflected by the guichroic mirror 7.8 and the half mirror 9 for each wavelength region depending on the properties of the sample particles, respectively, and are focused. optical lens 10.12
．． The light enters the photodetector 11.13°15 through the beam 14, and information mainly on the shape of the sample particle is obtained.

The pulse signal generated by the scattered light of the sample particles can be monitored by the monitor section 26, and the particle counting section 27 further counts the number of sample particles, so that the measurement can be performed once the data collection of the scheduled number of particles is completed. It is set to end automatically.

In this embodiment, four types of optical signals can be detected simultaneously from one particle, and the forward scattered light photodetector 5
The side-scattered light photodetectors 11, 13, and 15 photoelectrically convert the signals, which are input to amplifiers 16 and 17 to 19, and output to the analog processing section 20 after amplification. The amplified signal input to the anacog processing unit 20 is subjected to calculations such as peak detection, time integration, and logarithmic amplification.
2, the = signal is output to the first interface section 21.

In the first interface section 21, the analog processing section 20
A/D f copies the calculated signal from the communication line f.
The data is output to the microcomputer section 22 via. The signal inputted to the microcomputer section 22 is inputted to the microcomputer body 24 via the second interface section 23 having a data storage section, where calculations are performed.
The results are displayed on the display 25 as graphs such as histograms and cytograms.

FIG. 2 is a configuration diagram of the first interface section 21,
This interface section 21 is an analog processing section into which signals from the analog processing section 20 are input.

Tongue part 31, A/D converter 32 connected to Sui 2,000 part 31, / Hera 2 part 33 connected to A/D converter 32
．． While receiving a signal from the buffer section 33, the switch section 31. It is composed of an A/D converter 32 and a timing control circuit 34 that issues commands to the 8727 section 33.

The timing control circuit 34 is the analog processing section 20
After the specimen particles have passed, a transfer strobe signal a with a width of about 30 tiles is output by a trigger signal generated by the scattered light by the specimen particles. Microcomputer section 2 in Figure 1
2 is output. Then, during the processing of one sample particle,
The analog processing section 20 is provided with a cancellation circuit (not shown) that does not capture the signal even if the next sample particle passes through. The analog input unit 31 receives input from the analog processing unit 20 at once based on the outputs of the amplifiers 16 to 19 while the sample particles pass through the distribution unit 1a once and the next sample particles to be measured pass through. 4 types of analog signals A
, B, C, and D are sequentially switched by the IJ)19 signal from the timing control circuit 34 and output to the A7Dq converter 32. The A/D converter 32 is activated by the A/D conversion start signal C from the timing control circuit 34.
The input signal from the analog converter section 31 is A/D converted, and this A/D converted signal is output to the output converter section 33. At the same time as the A/D converter 32 outputs the conversion end signal d, the digital signal e is also output, and the transfer strobe signal a, the switching signal b, the conversion start signal C, the conversion end signal d, and the conversion end signal d are output. The respective timings of the signal e are as shown in FIG.

Converted data A', B', C of the converted signal e shown in FIG.
', D' are digital signals corresponding to the respective analog signals A, B, and C1D, and the shaded areas indicate unrelated data. These conversion signals e and change 4! ! ! ! end signal d
is outputted from the A/D converter 32 to the communication line f via the output buffer section 33. In this embodiment, a 12-bit A/D converter 32 is used, and the conversion signal e in FIG. 2 is a 12-bit data bus, and the signal g
is a signal indicating reception of data sent to the timing control circuit 34 via the microcomputer section 22 or 4227 section 33.

Within about 30 seconds after the sample particle passes, the photoelectrically converted signal undergoes analog calculation and is A/D converted by the first interface section 21, and at the same time, the converted signal e is sent to the microcomputer section 22. Second interface section 2
3 will be output.

FIG. 4 is a configuration diagram of the second interface section 23 of the microcomputer section 22. As shown in FIG. This second interface section 23 is provided with a buffer section 35 as an input section of the communication line f connected to the first interface section 21, and latch circuits 36 to 36 connected in parallel to this buffer section 35 are provided. 39, and eight latch circuits 36
The timing control circuit 40 controls the circuits 1 to 39.

Data A' that has been A/D converted by the interface section 21
. the respective write pulses i, j,
Then, ρ is generated by the timing control circuit 40 based on the transfer strobe signal a and the conversion end signal C input from the first interface unit 21 via the 4227 unit 35.

FIG. 5 shows the timing at which data A", B', C', and D' are written into the respective latch circuits 36, 37, 38, and 39. The end of the transfer strobe signal a is at i, j, This means the end of writing of the pulse of ρ, and the microcomputer main body 24 confirms the end of writing via the communication line n, and the written data is sent to the memory unit via the system bus m to the microcomputer main body 24. After the transfer strobe signal a ends,
It takes about 10 seconds for the microcomputer body 24 to store the data A° to D' in the storage section, and the storage of the data A' to D' in the storage section is completed in 40g seconds from the time the sample particles pass through the sample. do. Note that the signal p is a data storage completion signal of the microcomputer main body 24 that is output to the interface section 21.

In this way, by providing the dedicated interface section 21.23, even if a commercially available microcomputer is used, data can be acquired within 40 JL seconds after recognition of sample particles, so data can be acquired at least 20,000 particles per second. Data analysis is possible at high speed. Also, data acquisition time is 40. Since it is extremely short (seconds), free time can be used for analysis processing, etc., and real-time processing of data about sample particles is possible.

In this embodiment, as shown in FIG.
A/D conversion was performed using the D converter 32, but if the number of A/D converters is four and each A/D converter performs A/D conversion of each analog signal, data collection can be further improved. The time can be shortened, and the microcomputer main body 24 can perform other tasks in its free time.

[Effects of the Invention] As explained above, the particle analysis device according to the present invention can analyze information obtained from scattered light of sample particles by providing a dedicated interface and using a microcomputer.
Collects data for each sample particle without requiring complicated equipment,
Furthermore, it has the advantage that real-time processing can be easily performed.

[Brief explanation of drawings]

The drawings show an embodiment of the particle analysis device according to the present invention, in which Fig. 1 is a block diagram of the optical system and signal processing system, Fig. 2 is a block diagram of the first interface section, and Fig. 3 is a block diagram of the first interface section. FIG. 4 is a configuration diagram of the second interface section, and FIG. 5 is a timing chart thereof. Symbol l is a flow cell, la is a flow section, 2 is a laser light source, 3 is an imaging lens, 4 is a condensing lens, 5 is a photodetector, 6
is a condensing lens, 7.8 is a micro ink mirror, 9 is a half mirror 110, 12°14 is a Toko lens, 11.1
3. 15 is a photodetector, 16 to 19 are amplifiers, 20 is an analog processing section, 21 is a first interface section, 22 is a microcomputer section, 23 is a second interface section, 24 is a microcomputer main body, 25 is a display, 2
6 is a monitor section, 27 is a particle counting section, 31 is an analog switch section, 32 is an A/D converter, 33.35 is a buffer section, 34.40 is a timing control circuit, 36 to 3
9 is a latch circuit. Patent applicant Canon Co., Ltd. Figure 2 Figure 3

Claims (1)

[Scope of Claims] 1. An irradiation optical system that irradiates a laser beam to sample particles flowing through a flow section in a flow cell, a photometric optical system that measures light scattered from the sample particles by the laser beam, and from the photometric optical system. and a signal processing system that analyzes the plurality of types of signals obtained, and the signal processing system analyzes the plurality of types of analog signals that are simultaneously input each time one specimen particle passes,
A particle analysis device characterized in that the digital signal is converted into a digital signal by A/D conversion, and is analyzed in a data analysis section using a microcomputer section having an arithmetic section and a storage section. 2. The particle analysis device according to claim 1, wherein the signal processing system rearranges the obtained plural types of analog signals in time series and performs A/D conversion. 3. The signal processing system has an A/D converter corresponding to each of the plurality of types of analog signals, and simultaneously has an A/D converter.
A particle analysis device according to claim 1, which performs conversion. 4. The signal processing system includes an analog processing section that performs analog calculations on one or more types of optical signals obtained from one particle, and A/D conversion of the output signal from the analog processing section; The particle analysis device according to claim 1, further comprising an interface section for outputting to a microcomputer section. 5. The microcomputer section comprises an interface section having a storage section that temporarily stores information from the signal processing system, and a microcomputer body and display section that perform data analysis. The particle analysis device described in .