JPH02213745A - Device for inspecting sample - Google Patents

Abstract

PURPOSE:To increase a fluid velocity in the delivery except the measuring time and in cleaning and to reduce the time except the measuring time by providing a control means for changing the pressing force when the sample is inspected from that when the sample is not inspected. CONSTITUTION:When measurement is started, a sheathing liq. is supplied to a flow cell 19 by a sheathing liq. pump 1 to form a laminar flow of the sheathing liq. in the flow cell 19. A syringe is simultaneously moved at a high speed by a sample pump 3 to exert high pressure, and the sheathing liq. in a sample loop 16 is rapidly delivered. The air layer reaches a sensor 24, and the output signal is sent to a grain analysis circuit 35 and a liq. supply means control circuit 36. The circuit 35 begins to fetch data from the signals of the sensor 24 when the flow of the sample is stabilized in a flow cell part after a fixed time. When fetching is finished and recording is started, a signal is sent to the circuit 36 from the circuit 35, the syringe is moved at a high speed by the pump 3 to highly pressurize the inside of the loop 16, the sheathing liq. having a cleaning function is rapidly delivered, and a tube 18 and the flow cell 19 are quickly cleaned.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention relates to a sample testing device such as a flow cytometer that supplies a sample to a testing section and tests the sample at the testing section. ] In conventional sample testing devices, the operator inserts a sample tube into a container containing sample liquid, applies suction force to the sample tube, and then aspirates a specified amount of sample liquid, accumulates it in the device, and drains the water channel. After switching, the accumulated sample liquid was extruded by the extrusion sheath liquid to supply the sample liquid to the test area. The sample was then measured at the test site using optical, electrical, or electromagnetic measuring means. By using a sheath liquid with a cleaning function as the extrusion liquid to extrude the sample liquid,
The inside of the sample tube can be washed at the same time as pushing out the sample liquid.

[Problems to be Solved by the Invention] However, the conventional sample testing device described above requires a large amount of cleaning liquid during cleaning, and also requires a long cleaning time. As a result, the cleaning time occupies a large proportion of the measurement operation time, and when a plurality of samples are measured continuously, the total measurement time becomes long.

[Object of the Invention] The present invention aims to increase the fluid velocity during feeding and cleaning other than when measuring, thereby shortening the time other than when measuring, thereby improving throughput.

[Means for Achieving the Object] To achieve the above-mentioned object, the present invention provides a sample testing device that presses a sample liquid with an extrusion liquid, sequentially supplies the sample liquid to a test area, and tests the sample in the test area. The sample testing device is characterized by comprising a control means for changing the pressing force between when testing the sample and when not testing the sample.

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

FIG. 1 is a block diagram of an embodiment in which the present invention is applied to a flow cytometer. In the figure, 1 is a sheath pump, 2 is a washing pump, and 3 is a sample pump, each of which is composed of a syringe driven by a motor. These pumps! ,
2.3, the three-way valve 6.7.8, and the six-way valve 15, which will be described later, are controlled by a control circuit 36. A tube 9 is connected to the sheath pump 1, and the other end is connected to a sheath liquid inlet of a flow cell 19. A three-way valve 6 is provided in the middle of the tube 9 to which the tube 12 is connected, and the other end of the tube 12 is immersed in the sheath liquid 26 accumulated in the sheath liquid container 4. This sheath liquid 26 includes a cleaning liquid,
It has the effect of cleaning the inside of waterways. Tubes 10 and 11 are connected to the washing pump 2 and sample pump 3, respectively, and the other end of each tube is connected to a six-way valve 1.
5. In the middle of tubes 10 and 11,
A three-way valve 13.14 is provided, each connected with a tube 13.14, the other end of which is connected to the sheath liquid 2 in the sheath liquid container 4 having a cleaning function.
It is immersed in 6. The six-way valve 15 also includes a sample suction tube 17 and a sample liquid 27a that has been sucked.
A tube 18 is connected to both ends of the sample loop 16 for storing sample sample 27a to send the sample sample inlet 27a to the sample inlet of the flow cell 19. The sample loop 16 and tube 18 are made of transparent material, and a gas detector inside the tube is located near the six-way valve 15 on the side of the sample loop 16 connected to the sample suction tube 17 and near the flow cell 17 of the tube 18. Sensor 23
and 24 are provided. The output signals of these sensors are transmitted to a particle analysis circuit 35 and a control circuit 36.
It is connected to the.

FIG. 2 is a diagram showing details of the sensor 23, 24, which is composed of an LED 2B, a slit 29, and a photoelectric element 30 with a transparent tube 16 or 18 in between. In this configuration, since the degree of light transmission changes depending on the difference in the refractive index of the substance inside the tube, the sensor section can determine whether the inside of the tube is air or sample liquid.

A laser light source 38 is placed toward the test area in the flow cell 19, and a sample liquid such as a blood sample is wrapped in a sheath liquid and converged into a thin flow, causing the sample to pass through the test area one by one. Laser light emitted from the laser light source 38 is irradiated onto microparticles such as blood cells in the liquid, and light scattering occurs due to the microparticles. Direct light is removed by a stopper 39 placed opposite the laser light source 38 across the flow cell I9, and forward scattered light collected by a lens 40 located behind the stopper 39 is photometered by a photodetector 41. be done. The output of the photodetector 41 is sent to the particle analysis circuit 35
connected to. Although not shown in the drawings, an optical system for measuring side scattered light and fluorescence is provided in a direction perpendicular to the plane of the paper. These photometric outputs are also connected to the particle analysis circuit 35.

The measured waste liquid that has passed through the flow cell 19 is discarded into the waste liquid container 5 through a waste liquid tube 20 connected from the flow cell 19 to the waste liquid container 5.

Further, 25 is a test tube containing a sample liquid 27a, and the presence or absence of the test tube 25 is monitored by a container detection means 37. 2
Reference numeral 1 denotes a washing waste liquid receiver, in which the washing waste liquid that has been backwashed through the sample suction tube 17 is guided to the waste liquid container 5 through a waste liquid tube 22 .

Next, the operation process of sample supply in the above configuration will be explained.

In the initial state, each pump], 2.3 and three-way valve 6.
7.8, tubes 9.10, 11.12.13.14.
17, 18, the six-way valve 15, and the sample loop 16 are all filled with sheath fluid.

In the sample suction process, first, the six-way valve 16
At the position shown in the figure, the sample pump 3 performs a suction operation and stands by while sucking a small amount of air from the tip of the suction tube 17. Next, as shown in FIG. 4, the operator carries the sample liquid container 25 containing the sample liquid 27a, such as a blood sample or latex agglutination suspension, to the position of the suction tube 17, which is installed in the main body of the apparatus. When the user presses the suction start button, the sample pump performs a suction operation, and after a certain amount of sample liquid has been suctioned into the suction tube 17, the operation stops. Here, the small amount of air sucked earlier is the air layer 31a
This serves to separate the sheath liquid and sample liquid.

In the unlikely event that the sample liquid 27a runs out during suction, or if the tip of the suction tube 17 accidentally protrudes above the sample liquid level and sucks in air, the sensor 23 detects air and sends a signal. Liquid means] is sent to the control circuit 36, and the suction operation is stopped. Note that the suction operation is not stopped in the first air layer 31a passing through the sensor 23.

When suction of a certain amount of sample liquid from the sample liquid container 25 is completed and the operator removes the sample liquid container 25, the container detection means 37 sends a signal to the liquid feeding means] control circuit 36, and the sample pump 3 sends a signal to the suction tube 17. Start suction again to send the remaining sample liquid into the sample loop. At this time, air is suctioned from the tip of the suction tube 17, and when the air reaches the position of the sensor 23 as shown in FIG. 5, the suction operation of the sample pump 3 is stopped in response to a signal from the sensor 23.

Next, the six-way valve 15 rotates in the direction of arrow A to
The waterway connections will change as shown in the diagram. Here, the sample pump 3 performs a certain amount of liquid feeding operation, and the previously aspirated sample liquid is pushed forward with the sheath liquid across the air layer 32a. In addition,
The air layer 32a existing between the sheath liquid and the sample liquid separates the sheath liquid and the sample liquid without mixing them.

At this time, at the same time, the cleaning pump 2 pushes out the sheath liquid by a liquid feeding operation, and causes the sheath liquid to flow backward through a part of the six-way valve 15 and the sample suction tube 17, thereby cleaning. The cleaning waste liquid is discharged into a waste liquid receiver 21, passes through a waste liquid tube 22, and is discarded into a waste liquid container 5.

When the sample pump 3 pushes the sample liquid by a certain amount during the season, the liquid feeding operation is stopped, and the cleaning by the liquid feeding operation of the cleaning pump 2 is also stopped.

Next, the six-way valve 15 is rotated in the direction of the opening in the figure, and the connection of the water channels is switched as shown in FIG. Then, the sample pump 3 sucks a small amount of air from the tip of the suction tube 17, waits, and can move on to the next sample liquid suction process. Thereafter, by repeating the same process, multiple types of sample liquids are suctioned one after another, and as shown in FIG. 31a.

They are stored in order while being separated by 32a, 31b, 32b, 31c, and 32c.

Note that the number of types of sample liquid that can be aspirated and accumulated at one time is determined by the amount of sample liquid to be aspirated, the amount of sheath liquid, and the volume of the sample loop 16, that is, the inner diameter and length of the sample loop 16. Ru. The number of times the sample is aspirated is counted by the liquid feeding means control circuit 36, and the aspiration operation is controlled not to be performed when a predetermined number of times is reached.

The material of the sample loop 16 is a fluororesin that easily repels liquid, and the trailing end of the sample liquid flow is cut off by an air layer, so that almost no sample liquid remains on the inner wall of the sample tube after the sample liquid has passed, and the sample The cleaning effect is high even if the amount of cleaning liquid that flows after the liquid is small.

When multiple types of sample liquids have been accumulated in the sample loop as described above, the system enters a standby state for starting measurement as shown in Figure 8, and when the operator presses the measurement start button provided on the main body of the device, the sample Liquid measurement begins. Note that even if the limit number of sample liquids is not accumulated in the sample loop 16, the measurement can be started by pressing the measurement start button.

When the measurement start button is pressed, first the sheath pump 1
Send Season Night to Flow Cell 19? A laminar flow of the sheath liquid is formed in the flow cell 19 overnight.

At the same time, the sample pump 3 pumps the syringe at high speed to apply high pressure and rapidly transport the sheath liquid within the sample loop 16. The purpose of fast forwarding is to allow the sheath liquid unrelated to the measurement to pass quickly, thereby shortening the total measurement time as much as possible. As shown in FIG. 8, the air layer 31a
4 parts, and its output signal is sent to a particle analysis circuit 35 and a control circuit 36. In response to the signal from the sensor 24, the control circuit 36 causes the sample pump 3 to move the syringe at a normal speed and apply appropriate pressure to the sample loop so that the sample has an appropriate flow diameter in the flow cell section.

Further, the particle analysis circuit 35 starts acquiring data after a certain period of time has elapsed since the signal from the sensor 24, and after the flow of the sample becomes stable in the flow cell section.

Normally, data acquisition is completed after a set number of particles or time, but during data acquisition, an air layer 32 between the sample liquid 27a and the sheath liquid that pushes it out
When a passes through the sensor 24 section, the signal is sent to the particle analysis circuit 35 and the data acquisition is forcibly terminated.

When the data capture is completed, recording of the captured data on the recording medium is started, and at the same time, a signal is sent from the particle analysis circuit 35 to the control circuit 36, which causes the sample pump 3 to move the syringe at high speed to loop the sample. The tube 18 and flow cell 19 are quickly cleaned by pressurizing the inside of the tube 16 at a high level and rapidly transporting a sheath liquid having a cleaning function. The air layer 31b in the sample loop is the sensor 2
When the amount reaches 4 parts, the sample pump 3 is stopped and the liquid feeding operation is stopped. When the recording of the captured data that is being carried out at the same time is completed β, a signal is sent from the particle analysis circuit 35) to the control circuit 36, and the sample pump 3 starts the next sample liquid at the normal feeding speed. Resume liquid feeding.

By repeating the above steps, multiple types of sample liquids are automatically measured one after another. When the data acquisition of the last sample liquid 27c is completed, a signal is sent from the particle analysis circuit 35 to the liquid supply control circuit 36, the sheath liquid is pushed out by the sample pump 3, and the apparatus is stopped after the water channel is cleaned.

Various methods are well known for performing particle analysis using histograms and cytodulum processing based on the measurement data obtained in this way, and the calculations are performed in the particle analysis circuit 35.

Although the embodiments in which the present invention is applied to a flow cytometer have been described above, the present invention is not limited to this, and can be applied to a particle counter, a Coulter measuring instrument that measures particulates from the electrical impedance of a test part, Alternatively, the present invention can be widely applied to devices that sequentially supply samples to a test area, such as a particle measuring device using optical sound learning.

[Effects of the Invention] According to the present invention, the cleaning time and the feeding time can be shortened by increasing the feeding speed of the extrusion liquid during times other than during sample measurement. This shortens the measurement cycle when continuously and repeatedly measuring a plurality of samples, thereby increasing the throughput per unit of time.

[Brief explanation of the drawing]

1 and 3 to 8 are configuration diagrams of an embodiment of the present invention, FIG. 2 is a detailed diagram of a sensor for gas detection, and FIG. 9 is a detailed diagram of a sample loop. The main symbols in the diagram are: 1... Sheath pump, 2... Washing pump, 3... Sample pump, 4... Sheath liquid container 5... Waste liquid container, 6 .7.8...Three-way valve, 15...Six-way valve, 16...Sample loop, 23.24...Sensor, 25...Sample container, 38...・Laser light source, 41... photodetector,

Claims (1)

[Scope of Claims] 1. In a sample testing device that presses a sample liquid with an extrusion liquid and sequentially supplies it to a test area, and tests the sample in the test area, the pressing force is increased during sample testing and during non-testing. A sample inspection device characterized by comprising a control means for changing. 2. The sample testing device according to claim 1, wherein the discrimination between sample testing and non-testing is performed by detecting a boundary between the sample liquid and the extrusion liquid at a predetermined position. 3. The sample testing device according to claim 2, wherein a gas is interposed at the boundary between the sample liquid and the extrusion liquid. 4. The sample inspection device according to claim 3, wherein the detection means is an optical sensor that distinguishes between gas and liquid in the supply path. 5. The sample inspection device according to claim 1, wherein the supply route is cleaned by the extrusion liquid. 6. Claim 1, wherein the sample is inspected by irradiating the flowing sample with light and detecting the optical reaction in the test section.
Sample testing equipment as described.