JPH0210134A - Flow type grain analyzer - Google Patents

Abstract

PURPOSE:To increase a flow velocity of a washing liquid for flowing in a sheath flow forming chamber and to improve the eliminating capacity for a bubble and a fragment of a grain, etc. by providing a washing liquid outflow port on the sheath flow forming chamber. CONSTITUTION:When a plunger 19 is moved upward as soon as a sample is sucked, and a sheath liquid is fed to a sheath flow forming chamber 4, the sheath liquid flows out of an air vent exclusive port 4C, discharged into a washing tank 15 which can be observed visually from a port 15b, and the inside of the sheath flow forming chamber 4 is washed. Subsequently, the sample measurement is executed, and also, the outside periphery of a sample suction nozzle 9 is washed, a washing liquid drops into the washing tank 15, and next, when a plunger 18' is allowed to ascend, and the sheath liquid is fed to the sheath flow forming chamber 4 from a sheath liquid inflow port 4a, the sheath liquid flows in reverse in a sample feed liquid tube 16 from a sample outflow port 4b, discharged to the washing tank 15 and washing is executed.

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application This invention relates to a flow-type particle analyzer applied to cell analysis, etc., and more specifically, to automatic cleaning thereof.

(b) Conventional technology A flow-type particle analyzer allows a sample containing particles to be analyzed to flow together with a sheath liquid through a flow channel with a small cross-sectional area, and hydrodynamically focuses the sample on the central axis of the flow channel. The particles are made to flow in a line, and the particles are irradiated with a laser beam to analyze the shape, type, etc. of the particles based on the scattered light or fluorescence (hereinafter referred to as signal light) generated by the particles.

2. Description of the Related Art Conventional flow-type particle analyzers are known to have a configuration in which sample suction and flow cell cleaning can be performed automatically for the purpose of efficiently performing particle analysis. The flow cell includes the flow channel and a sheath flow forming chamber provided upstream of the flow channel. The sheath flow forming chamber is provided with a sheath liquid inlet and a sample inlet that are connected to the liquid feeding system.

A sheath liquid sent by a liquid sending means such as a syringe flows into the sheath liquid inlet. On the other hand, from the sample inlet,
The sample that has been sucked into the sample liquid feeding path from the sample suction nozzle is pushed out and flows into the sample liquid feeding path.

In this way, the particles in the sample flow in a line on the central axis of the flow channel, and the particles are analyzed.

Immediately after (or just before) the analysis, the flow cell, sample suction nozzle, and sample liquid feeding path are cleaned.

To clean the sample suction nozzle, send the cleaning liquid to the sample suction nozzle, let this cleaning liquid flow out from the sample suction nozzle,
Wash away any remaining sample.

Further, to clean the flow cell and the sample liquid feeding path, a cleaning liquid is sent to the sample liquid feeding path, and the sample remaining in the sample liquid feeding path is washed away into the sheath flow forming chamber. Then, the cleaning liquid flows into the sheath flow formation chamber from the sheath liquid inlet, and this cleaning liquid is poured into the flow channel together with the cleaning liquid (containing excess sample) flowing in from the sample inflow path. The flow cell is washed by flushing.

(c) Problems to be Solved by the Invention In the conventional flow-type particle analyzer described above, when cleaning the sample suction bow nozzle, only the inner periphery of the sample suction nozzle is cleaned. For this reason, there is a problem in that the sample adhering to the outer periphery of the sample suction nozzle mixes with other samples during the next analysis, reducing the reliability of the analysis.

In addition, when cleaning the flow cell, the cleaning liquid that flows in from the sheath liquid inlet flows through a flow channel with a small cross-sectional area and is discharged to the outside, making it impossible to obtain a sufficient flow rate and forming a sheath flow. There was a problem in that it was difficult to wash away the bubbles and particle fragments in the chamber, and the bubbles and particle fragments adhered to the sheath flow forming chamber, causing disturbance of the sheath flow.

Furthermore, as the cleaning liquid containing excess sample flows through the sample inlet and flows through the flow channel, particles and chemical substances (proteins, etc.) contained in the sample adhere to the inner wall of the flow channel, and are transferred to the laser beam particles. There was a problem that the irradiation efficiency was reduced.

On the other hand, the liquid discharged from the flow channel and other liquids from the liquid delivery system are led directly to the outlet of the device, so it is not possible to check the discharge status of the liquid from each part. It is not easy to monitor the operating status of the system, and it is especially difficult to discover the cause of a failure.

This invention was made in view of the above, and is a flow-type particle analyzer that has high analysis reliability, stable sheath flow, high efficiency of laser irradiation to particles, and easy detection of the cause of failure. intended to provide.

(d) Means for Solving the Problems In order to solve the above problems, the flow-type particle analyzer of the present invention includes a sheath flow forming chamber in which a sample inlet and a sheath liquid inlet are provided, and a flow channel. a flow cell, a sheath liquid sending means for sending the sheath liquid to the sheath liquid inlet, a sample suction liquid sending means for sending the sample sucked into the sample liquid feeding path from the sample suction nozzle to the sample inlet, and the flow channel. a light beam irradiation means for irradiating a light beam onto particles flowing in a line; a photodetector for detecting signal light from the particles irradiated with the light beam; analysis processing means for performing an analysis process, comprising: a washing liquid outlet provided in the sheath flow forming chamber; and a washing liquid flowing back from the sample inlet into the sample liquid feeding path; a backflow cleaning means for cleaning the cleaning liquid, and a drainage concentration section that concentrates the drainage flowing out from the cleaning liquid outlet, the drainage from the backflow cleaning means, and the drainage from the flow channel, and allows the state of each drainage to be visually observed. and a cleaning liquid nozzle that is integrally supported with the sample suction nozzle and that sprays cleaning liquid onto the outer periphery of the sample suction nozzle.

(e) Function In the flow type particle analyzer of the present invention, since the sheath flow formation chamber is provided with a cleaning liquid outflow port, the cleaning liquid that flows into the sheath flow formation chamber from the sheath liquid inlet flows through the flow channel and not through the flow channel. Since the cleaning liquid flows out from the wide-channeled cleaning liquid outlet, the flow rate of the cleaning liquid flowing inside the sheath flow forming chamber can be increased, and bubbles, particle fragments, etc. inside the sheath flow forming chamber can be removed better than before.

In addition, since the cleaning liquid in the sheath flow formation chamber flows back from the sample inlet to the sample wave channel and washes away the sample remaining in the sample channel, excess sample does not flow through the flow channel. Particles and chemical substances in the sample are less likely to adhere to the inner walls of the flow channel.

Furthermore, since the condition of each drained liquid can be visually observed, if a failure occurs in any part of the liquid delivery system, the location can be quickly identified.

In addition, since the outer periphery of the sample suction nozzle is also cleaned, the next sample will not be contaminated with the previous sample, making it possible to improve the reliability of analysis.

(F) Embodiment An embodiment of the present invention will be described below with reference to FIGS. 1 and 2.

In this embodiment, the present invention is applied to a cell analysis device, and FIG. 1 is a diagram illustrating the structure of a cell analysis bag W1 of the embodiment.

2 is a flow cell, which is composed of a flow channel 3 and a sheath flow forming chamber 4.

The flow channel 3 is made of a material transparent to light having the wavelength of the laser 5 used, such as synthetic quartz. The sheath flow forming chamber 4 is equipped with a sheath liquid inlet 4a, a sample inlet 4b, and a bubble removal exclusive port (cleaning liquid outlet) 4c.

Around the flow channel 3 are a laser 5 and a photodetector 6.
is placed. A laser beam from the laser 5 is irradiated onto cells flowing in a line within the flow channel 3 due to hydrodynamic focusing effects. The signal light of this cell is
The light is received by the photodetector 6 and converted into an electrical signal. This electrical signal is processed by a microcomputer (analysis processing means) not shown, and the cells are analyzed. Note that this invention does not involve analysis processing as a main part, so the details will be omitted.

7 is a sample suction section. Reference numeral 8 denotes a holder that can swing around a fulcrum 8a, and a sample suction nozzle 9 and a cleaning liquid jet nozzle 10 are integrally attached to the tip. 11 is a drive motor for the holder 10, and this drive motor 11
The roller 1 at the tip of the crank 12 is attached to the rotating shaft of the
3 comes into contact with the lower surface of the holder 8. The holder 8 swings as the drive motor 11 rotates, and the test suction nozzle 9 and the cleaning liquid jet nozzle 10 move up and down. Note that 14 is a sample container in which the sample S is placed.

Reference numeral 15 denotes a cleaning tank (effluent concentration section), which is made of a visually visible material such as transparent or smoked resin.

The upper part of this cleaning tank 15 is open, and when the holder 8 moves downward, the sample suction nozzle 9 and the cleaning liquid jetting nozzle 1 are opened.
0 is located above. This cleaning tank 15 is connected to the drain port 1
5a, 15b, and 15c, and these drain ports 15a, 15b, and 15c are arranged so that it can be seen from which boat the drain is flowing out. The drain port 15a is connected to the flow channel outlet 3a via a two-way valve v1, and the drain port 15b is connected to the bubble removal exclusive port 4c via a two-way valve V2.

Further, the drain port 15c is connected to the base end side of the sample suction tube 16 via the two-way valve 4.

On the other hand, the waste liquid concentrated in the waste liquid concentration section 15 is led to an outlet (not shown) of the cell analyzer via a valve V3.

17, 18, and 19 are syringes (sheath liquid feeding means, sample suction liquid feeding means), respectively. Each cylinder plunger 17', 18', 19'' has a slider 2.
0 is attached, and each slider 20 is screwed into a ball screw 21.

The ball screw 21 is coupled to a rotating shaft of a motor 23 via a coupling 22 . The slider 20 moves due to the rotation of the motor 23, and as a result, each plunger 17'',
18' and 19' are respectively driven up and down in the plane of FIG. Note that each motor 23 is controlled by a control circuit (not shown).

The syringe 17 is connected to the proximal end of the sample suction tube 16. On the other hand, the distal end of the sample suction tube 16 is connected to the port C of the three-way valve vT, and the sample inlet 4b is connected to the port C of the three-way valve vT.
, sample suction tube 9.

The syringe 18 is connected to the base end side of the sample liquid feeding tube 16 via a two-way valve 5, and is connected to the cleaning liquid jet nozzle 10 via the two-way valve 6, and to the sheath liquid inlet 4a via the two-way valve V7. Send the sheath fluid to each.

The syringe 19 also sends the sheath liquid to the sheath liquid inlet 4a via the three-way valve (2).

24 is a sheath liquid suction nozzle, and a sheath bottle 25
Suction up the sheath fluid inside. The sheath liquid sucked up by the sheath liquid suction nozzle 24 is supplied to the syringe 19 via the three-way valve V1o, and to the syringe 18.17 via the two-way valves V, , V, .

Two-way valve v1, v2, V3, ■4, ■5, V6, ■7, v
8, ■7, ■Io, and three-way valve ■7 are each electromagnetic valves, and are controlled by the control circuit. Of course, each of the valves (1) to VIO1 (2) is not limited to a solenoid valve.

Next, the operation of the cell analyzer according to the embodiment will be explained with reference to FIG. In addition, two-way valve ■1, ■2, ..., VIO
If there is no explanation, the two-way valve is closed.

In the starting state, the holder 8 is in an upwardly swung state, and the sample container I4 is set as shown in FIG. 1, and the lower end of the sample suction nozzle is located in the sample S. First, the sample S is aspirated [step (hereinafter referred to as ST) 1
].

In STI, the three-way valve ■□ is set to the A side, the two-way valve v5 is opened, the plunger 18'' is moved downward, and the sample S is sucked into the sample liquid feeding tube 16 through the sample suction nozzle 9.

Meanwhile, simultaneously with the sample suction, the sheath flow formation chamber is cleaned (Sr1). In this Sr1, two-way valve V2, ■
, and move the plunger 19 of the syringe 19 upward.
The sheath liquid in the syringe 19 is sent to the sheath flow forming chamber 4. The sheath liquid that has flowed into the sheath flow forming chamber 4 flows out from the bubble removal port 4C and flows into the cleaning tank 1 from the port 15b.
It is discharged within 5 days. At this time, the sheath liquid washes away bubbles, cell debris, etc. in the sheath flow forming chamber 4.

Next, sample measurement (Sr1) is performed, and in parallel with this, sheath fluid is supplied to the syringe 18 (Sr1) and outer periphery of the sample suction nozzle 9 is cleaned (Sr1).

For Sr1, switch the three-way valve ■1 to the B side, and open the syringe 17.
Raise the plunger 17゛ of the sample liquid supply tube 1.
6 is sent to the sample inlet 4b. At the same time, the two-way valve V1, ■ is opened, and the plunger 19 of the syringe 19
'' rises, and the sheath liquid is sent to the sheath liquid inlet 4a.

A sheath flow is formed within the flow channel 3, and the cells flow in a line due to the hydrodynamic focusing effect. These cells are irradiated with a laser beam from a laser 5, a signal light is received by a photodetector 6, and analysis is performed in the same manner as in the past. The waste liquid discharged from the flow channel 3 is discharged into the cleaning tank 15 from the port 15a.

At Sr1, the two-way valve 8 is opened, the plunger 18' of the syringe 18 is lowered, and the sheath liquid sucked from the sheath liquid suction nozzle 24 is supplied to the syringe 18.

Continuing from Sr1 In Sr1, the holder 8 is first moved to the position 8'' shown by the broken line in FIG. Then, the three-way valve 6 is opened, the plunger 18' is raised, and the sheath liquid in the syringe 18 is ejected from the cleaning liquid jetting nozzle 10, thereby cleaning the outer circumference of the sample suction nozzle 9. The sheath liquid used for this cleaning falls into the cleaning tank 15.

Next, the sample liquid feeding tube 16 and the sample inlet 4b are cleaned (Sr1). This cleaning is performed using two-way valves V, ,V,
Open , leave ■ on the B side, and press plunger 18.
The sheath liquid in the syringe 18 is sent to the sheath flow forming chamber 4 through the sheath liquid inlet 4a. The sheath liquid that has flowed into the sheath flow forming chamber 4 flows through the sample outlet 4b.
, the sample liquid flows backward in the sample liquid feeding tube 16 and flows through the port 15.
c and is discharged into the cleaning tank 15 for cleaning.

When Sr1 is completed, internal cleaning of the sample suction nozzle 9 (Sr1 is completed)
T7a) and sheath fluid supply to the syringe 17 (ST7b
) are performed in parallel. In Sr1 a, the three-way valve 7 is switched to the A side, the three-way valve 5 is opened, the plunger 18 is raised, and the 1-day-old sheath fluid is aspirated into the syringe through the sample liquid supply tube 16. The sample suction nozzle 9 is cleaned by flowing out from the nozzle 9. On the other hand, Sr1
At step b, the plunger 17' is lowered and the sheath fluid is supplied into the syringe 17.

In parallel with the processing of Sr1 and 5T7a/7b, liquid is being supplied to the syringe 19 (Sr1). This liquid supply is performed by opening the three-way valve V1o and lowering the plunger 19''.

When the processing of 5T7a/7b and Sr1 is completed, move the holder 8 to the upward movement position (solid line position shown in Figure 1).
9), Finish the measurement for this sample.

In the processing of Sr1, Sr3, and Sr1, port 1
5a, 15b, and 15c, it is easy to check whether each process is being performed normally, and even if an abnormality occurs, each port 15a,
The location where the abnormality has occurred can be easily discovered based on the presence or absence of drainage from 15b and 15c.

In the above embodiment, the sheath liquid is also used as the cleaning liquid.
The cleaning liquid may be separate from the sheath liquid, and the design can be changed as appropriate.

Further, in the above embodiment, the holder 8 is configured to swing, but when the holder 8 is in the upward movement position shown by the solid line in FIG.
It has the advantage that it can be brought to the suction position more easily than diagonally downward. Of course, the sample suction nozzle may be moved vertically up and down.

(G) As described in detail of the invention, the flow-type particle analyzer of this invention has the following features:
A cleaning liquid outlet provided in the sheath flow formation chamber, a backflow cleaning means for causing the cleaning liquid to flow back into the sample liquid passage from the sample inlet of the sheath flow formation chamber to clean the inside of the sample liquid passage, and a cleaning liquid outlet provided in the sheath flow formation chamber. Concentrating the outflowing liquid, the liquid draining from the backwashing means, and the liquid draining from the flow channel,
The device is characterized by comprising a drain concentration part where the state of each drain liquid can be visually checked, and a cleaning liquid nozzle that is supported integrally with the sample suction nozzle and sprays cleaning liquid onto the outer periphery of the sample suction nozzle. This has the advantage that the ability to wash out air bubbles, particle fragments, etc. in the sheath flow forming chamber is improved, and the sheath flow is less likely to be disturbed. Furthermore, since excess sample does not flow within the flow channel, particles and chemical substances in the sample do not adhere to the inner wall of the flow channel, which has the advantage of preventing a decrease in laser irradiation efficiency. Furthermore, there is a low possibility that the sample from the previous measurement adhering to the outer periphery of the sample suction nozzle will be mixed into the next sample, which has the advantage of improving the reliability of analysis. In addition, it has the advantage that the condition of each drainage liquid can be visually observed, making it easy to discover failures and investigate the location of failures.

[Brief explanation of the drawing]

Each of the drawings shows an embodiment of the present invention, and FIG. 1 is a diagram illustrating the configuration of a cell analysis device according to this embodiment, and FIG.
The figure is a flow diagram illustrating the operation of the cell analysis device. 2: Flow cell, 3: Flow channel 4 nitrogen flow formation chamber, 4a: Sheath liquid inlet 4b = sample inlet, 4c: Dedicated bubble removal port 5: Laser, 6: Photodetector 9: Sample suction nozzle, 10: Cleaning liquid jet nozzle 15: Cleaning tank, 16: Sample liquid feeding tube 17.1 day.1
9: Syringe.

Claims (1)

[Claims] (1) A flow cell composed of a sheath flow forming chamber and a flow channel in which a sample inlet and a sheath liquid inlet are provided, a sheath liquid feeding means for feeding the sheath liquid to the sheath liquid inlet, and a sample suction nozzle. A sample suction liquid feeding means for feeding the sample sucked into the sample liquid feeding path to the sample inlet; a light beam irradiation means for irradiating a light beam onto the particles flowing in a line in the flow channel; A flow type particle analyzer comprising a photodetector that detects signal light from irradiated particles, and an analysis processing means that performs analysis processing of particles based on the light reception signal of the photodetector, the sheath flow formation. a cleaning liquid outlet provided in the chamber; a backflow cleaning means for causing the cleaning liquid to flow back from the sample inlet into the sample liquid feeding path to clean the inside of the sample liquid feeding path; and a waste liquid flowing out from the cleaning liquid outlet and the backflow. A drainage concentration part that concentrates the drainage from the cleaning means and the drainage from the flow channel and allows the state of each drainage to be visually observed; and a drainage concentration part that is integrally supported with the sample suction nozzle, and the cleaning liquid is placed around the outer periphery of the sample suction nozzle. A flow-type particle analyzer characterized by comprising a cleaning liquid nozzle that sprays a cleaning liquid nozzle.