JPH01118747A - Particle analyzer - Google Patents

Abstract

PURPOSE:To set a flow diameter matching with fluid particles automatically by providing a particle diameter information detecting means which detects particle diameter information on the sample particles and a means which sets the flow diameter corresponding to the detect particle diameter. CONSTITUTION:A control arithmetic circuit 17 controls a regulator 3 for sheath liquid and a regulator 4 for sample liquid at the start of measurement to set the flow diameter of a flow of sample fluid in a flow cell 12 to a certain specific value. The sample particles which flow at this time are irradiated with laser light from a laser 19. There is a correlation between the quantity of forward scattered light and the particle diameter, so a photodetector 22 detects the quantity of the forward scattered light at this time and compares it with a light quantity-particle size conversion table in the memory of a circuit 17 to calculate the particle size. Then the circuit 17 controls the regulators 3 and 4 so as to obtain a flow diameter nearly equal to the found particle size. Here, the pressure values of sheath liquid Sh and sample liquid Sa for obtaining the specific flow diameter are predetermined and stored in the memory of the circuit 7. Thus, the flow diameter matching with the particle size is set automatically.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a particle analysis device, and particularly to a so-called flow cytometer that performs particle analysis by flowing a sample liquid containing specimen particles while being wrapped in a sheath liquid.

[Conventional technology] A flow cytometer applies a predetermined pressure difference between a sample liquid, which is a cell suspension solution, and a sheath liquid (e.g., physiological saline) on the outside, causes the sample liquid to flow at high speed, and converges the flow hydrodynamically. This device analyzes the properties, structure, etc. of the sample particles by irradiating the sample particles flowing one by one to the convergence position with a laser beam, detecting the scattered light and fluorescence from the sample particles with a photodetector, Used in fields such as cytology, hematology, oncology, and genetics.

The sample liquid and the sheath liquid are each stored in a container, and the air in the container is pressurized by a pressurizing system such as a compressor or a nitrogen gas cylinder, so that each liquid fluidly converges on the flow cell section, which is the measuring section. Note that the applicant's earlier application, Japanese Patent Application No. 62-185841, discloses a pressure control system, in which a pressure sensor is provided between the sheath liquid container and the flow cell, and the output of the pressure sensor is constantly feedback-controlled. The flow diameter of the sample liquid is stabilized by avoiding the influence of pressure fluctuations due to filter clogging or drop in sheath liquid.

The flow diameter of the sample liquid flowing inside the flow cell is determined in advance by considering the type of sample particles and the measurement speed, but the setting of this flow diameter is determined by the pressure of the air pressurized into the containers containing the sheath liquid and sample liquid. This is done by setting in advance a predetermined value.

[Problems to be Solved by the Invention] However, in the conventional example described above, when the type of sample particles is unknown, or when the size of the particles is unknown even if the type is known, the optimal flow diameter cannot be determined. First, accurate measurements were difficult because the sample particles drifted in a direction intersecting the flow direction.

[Means for Solving the Problems] As a means for solving the above-mentioned problems, for example, in this embodiment, sample particles are irradiated with a laser beam at a predetermined flow diameter at the start of measurement, and the output of the forward scattered light is calculated. The particle diameter is determined, and then the flow diameter is reset to correspond to the determined particle diameter, and the main measurement is performed.

[Example code] Embodiments of the present invention will be described in detail below with reference to the drawings.

In FIG. 1, 1 is a compressor which is a source of compressed air, and an air tube 2 connected to this compressor is branched into two branches. The branched air tubes 2 are connected to a sheath liquid container and an electric regulator 3.4 for pressure adjustment provided for the sample liquid, respectively.
It is connected to a sheath liquid solution 5 and a sample liquid container 6 that airtightly store the sheath liquid sh and sample liquid Sa. The sheath tube 7 immersed in the sheath liquid sh in the sheath liquid container 5 includes a filter 8 for removing impurities, a pressure sensor 9,
The sheath liquid is guided into the nozzle 11 via the sheath liquid inflow control valve 10. Further, the sample tube 13 immersed in the sample liquid a in the sample liquid container 6 is guided into the nozzle 11 via the sample liquid inflow control valve 14, and its tip is connected to the flow cell connected to the upper end of the nozzle 11. It is aimed at 12. The sample liquid and sheath liquid are caused to flow from the bottom to the top against gravity so that air bubbles that are unexpectedly generated in the flow cell 12 flow with the fluid and are quickly removed. A waste liquid tube 15 is connected to the upper end of the flow cell 12, and the other end is connected to a waste liquid container 16. The control calculation circuit 17 receives the output of the pressure sensor 9 manually and also receives the output of the scattered light detector. The output from this is input to the sample liquid regulator 4 and the sheath liquid regulator 3.

When the liquid level of the sheath liquid sh falls, or when the filter 8 is clogged and the output of the pressure sensor 9 falls below the reference value, the internal pressure of the container 5 is increased by the regulator 3, and the output of the pressure sensor 9 is increased. Pressure control is performed by matching the reference value. In addition, 19 is a measurement laser for particle analysis, 20 is a condensing lens, 21 is a light receiving lens,
22 is a photodetector, and a combination of a light receiving lens and a photodetector is set in the laser irradiation direction and on the side.

Next, a method for setting the flow diameter according to the particle diameter of the sample particles will be explained.

At the start of measurement, the control calculation circuit 17 controls the sheath liquid regulator 3 and the sample liquid regulator 4 to set the flow diameter of the sample liquid in the flow cell 12 to a certain predetermined value. At that time, the flowing sample particles are irradiated with laser light. Since there is a correlation between the amount of forward scattered light and the particle diameter, the amount of forward scattered light at this time is detected by the photodetector 22 and compared with the light amount-particle diameter conversion table in the memory of the control calculation circuit 17. By doing so, the particle size is calculated. The control calculation circuit 1 determines the flow diameter that approximately matches the determined particle diameter.
7 controls the sheath liquid and sample liquid regulators. Here, the pressurization values of the sheath liquid and sample liquid to obtain a predetermined flow diameter are determined in advance and stored in the memory of the control circuit 17. Note that the flow diameter is determined by the pressure ratio of the sheath liquid pressure value and the sample liquid pressure value, and the larger the pressure ratio is, the greater the sheath liquid pressure value relative to the sample liquid pressure value, the greater the flow diameter. becomes smaller. In addition, when the pressure values of both are increased, the flow rate becomes faster. After the above adjustments are completed, measurements for particle analysis are started.

In the above explanation, the particle diameter was calculated only from the output of forward scattered light, but the type of particle was determined from the output of forward scattered light and side scattered light, and the particle diameter was calculated by making corrections according to the properties of the particle. By calculating , a more accurate particle size can be obtained. Furthermore, depending on the particle, it is also possible to calculate the particle diameter from only the output of side scattered light.

Note that the present invention is also applicable to a so-called jet-in-air type flow cytometer, that is, a device that converges a flow in the air and irradiates a laser beam thereon to perform particle analysis.

[Effects] According to the present invention, even if the sample particles are unknown or the particle size of the sample particles is unknown, the flow diameter suitable for the sample particles can be automatically set, making it easy to handle. We provide particle analysis equipment with high measurement accuracy.

[Brief explanation of the drawing]

FIG. 1 is a diagram of an embodiment of the present invention. In the figure, 1 is a compressor, 2 is an air tube, 3 is an electric regulator for the sheath, 4 is an electric regulator for the sample, 5 is a sheath liquid container, 6 is a sample tube, 7 is a sheath tube, 8 is a filler, 9 is a pressure sensor, 10
1 is a sheath liquid inflow control valve, 11 is a nozzle, 12 is a flow cell, 13 is a sample tube, 14 is a sample liquid inflow control valve, 15 is a waste liquid tube, 16 is a waste liquid container, 17 is a control calculation circuit, 19 is a laser, and 20 is a 21 is a light-receiving lens, and 22 is a photodetector.

Claims (1)

[Claims] 1. A sheath liquid is wrapped around a sample liquid containing analyte particles, and a predetermined pressure difference is applied between the sample liquid and the sheath liquid to fluidically converge the sample liquid to a predetermined flow diameter. , a particle analysis device that performs particle analysis by irradiating the measurement light onto the convergence position and measuring scattered light or fluorescence scattered by the sample particles, comprising a particle size information detection means for detecting particle size information of the sample particles; 1. A particle analysis device comprising means for setting a flow diameter according to a particle diameter determined. 2. The particle analysis device according to claim 1, wherein the particle size information detecting means detects the particle size based on the output of scattered light in a forward direction with respect to the irradiation light direction among the scattered lights.