JPH02213746A - Device for inspecting sample - Google Patents

Abstract

PURPOSE:To carry out normal measurement even when air is introduced into a sample pipe in the supply of sample by providing a control means for stopping pressurization or suction in accordance with the output of a detecting means. CONSTITUTION:When a suction starting button is pushed, a sample pump 3 is driven, hence a prescribed amt. of a liq. sample 27a is sucked into a suction tube 17, and then the pump is stopped. A small amt. of air previously sucked form an air layer 31a, and a sheathing liq. and the liq sample are separated by the layer 31a. However, when the amt. of the liq. sample 27a prepared in a liq. sample container 25 is smaller than the prescribed amt. to be sucked, the liq. sample 27a is run out during the suction, the container 25 is emptied, and the air is sucked into the tube 17. In such a case, the air flowing in the tube 17 is optically detected by a sensor 23, the signal is sent to a liq. supply means circuit 36, and suction is immediately stopped. As a result, measurement is regularly carried out at all times.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a sample testing device such as a flow cytometer.

[Prior Art] Conventionally, in sample testing devices such as flow cytometers and particle counters, a sample tube is inserted into a sample liquid container containing the sample liquid to be measured, and then the sample tube is inserted into the sample liquid container. A sample supply method is known in which a specified amount of sample liquid is forced into a sample tube by sealing and fixing the container and pressurizing the inside of the container, and directly supplying the sample liquid to the test area.

Alternatively, the operator inserts the sample tube into the sample liquid container, applies suction to the sample tube, and
- There is a known sample supply method in which a specified amount of sample liquid is aspirated and accumulated in the device, the water channel is switched, and then the accumulated sample liquid is pressurized with a syringe or the like and the sample liquid is supplied to the test area. .

[Problems to be Solved by the Invention] However, in the conventional sample testing device described above, if the sample liquid prepared in the sample liquid container is less than a predetermined amount, all the sample liquid is sent. Even after the sample liquid container was emptied, pressurization or suction continued, causing air to be pumped into the sample tube.

In addition, in a sample inspection device using the former method,
If the tip of the sample tube protrudes above the surface of the sample liquid while pressurizing the sample, or in a sample testing device using the latter method, the operator may remove the sample liquid container from the sample tube while aspirating the sample. Air was also pumped into the sample tube when it was removed from the sample tube.

[Object of the Invention] An object of the present invention is to provide a sample testing device that does not adversely affect the measurement operation even if air is sent into the sample tube due to some kind of accident during sample supply.

[Means for Achieving the Object] The present invention achieves the above-mentioned object by supplying a sample liquid to a test area by pressurization or suction from a sample liquid container containing a sample liquid, and discharging the sample at the test site. A sample testing device for testing a sample, which is provided in the middle of a sample supply route and has a detection means for detecting gas in the sample supply route, and a control means for stopping pressurization or suction operation based on the output of the detection means. This is a sample inspection device characterized by:

[Example] Hereinafter, an example in which the present invention is applied to a flow cytometer will be described in detail using the drawings.

FIG. 1 is a configuration diagram of an embodiment. 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 1.2.3 and the three-way valve 6. described below.
7.8 and the six-way valve 15 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 a sheath liquid 26 having a cleaning function and stored in the sheath liquid container 4. 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 15. A three-way valve 13.14 is provided in the middle of the tubes 10, 11, and the tubes 13.14 are connected to each other.
The other end of the sheath liquid 26 is immersed in a sheath liquid 26 in a sheath liquid container 4 having a cleaning function. The six-way valve 15 includes
In addition, a sample suction tube 17 and a tube 18 for sending the aspirated sample liquid 27a to the sample inlet of the flow cell 19 are connected to both ends of the sample loop 16 for storing the aspirated sample liquid 27a. The sample loop 16 and the 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 2
3 and 24 are provided. The output signals of these sensors are transmitted to the particle analysis circuit 35 and the liquid feeding means] control circuit 3.
6.

FIG. 2 is a diagram showing details of the sensor 23, 24, which is composed of an LED 28, 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. The direct light is removed by a stopper 39 provided opposite the laser light source 38 with the flow cell 19 in between, and the 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.

Reference numeral 21 denotes a washing waste liquid receiver, in which the washing waste liquid that has backwashed 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 1.2.3 and three-way valve 6.
7.8, tube 9.10.11.12.13.14.
17, 8, six-way valve 15, and 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 brings 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, and supports it by hand. . When a suction start button provided on the main body of the apparatus is pressed, the sample pump performs a suction operation, and after a predetermined amount of sample liquid has been suctioned into the suction tube 17, the operation is stopped. The small amount of air sucked in here becomes an air layer 31a,
It plays the role of separating the sheath liquid and sample liquid.

However, if the amount of sample-ti 27a prepared in advance in the sample liquid container 25 is less than the specified amount to be aspirated, the sample liquid 27a will run out during suction and the inside of the sample liquid container 25 will become empty. Air is sucked into the suction tube 17. Alternatively, the operator may accidentally remove the sample liquid container 25 from the suction tube 1 during suction.
Even if it is separated from the suction tube 7, the tip of the suction tube 17 will come out above the sample liquid level and suck in air.

In these cases, the sensor 23 optically detects the air flowing into the suction tube, and a signal is sent to the liquid feeding means control circuit 36 to immediately stop the suction operation. 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 I&pull 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. At this point, the sample pump 3 pumps a certain amount of liquid, 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 liquid is pushed forward by a certain amount with the sheath liquid by the sample pump 3, the liquid feeding operation is stopped, and the cleaning by the liquid feeding operation of the cleaning pump 2 is also stopped.

As described above, when the sample liquid has been accumulated in the sample lube, it enters the measurement start standby state, and when the operator presses the measurement start button provided on the main body of the device,
Measurement of the sample liquid begins.

When the measurement start button is pressed, first the sheath pump 1
sends the sheath liquid to the flow cell 19
A laminar flow of sheath liquid is formed within the sheath.

At the same time, the sample pump 3 pumps the syringe at high speed to rapidly transport the sheath liquid within the sample loop 16. The purpose of fast forwarding is to allow the sheath liquid unrelated to measurement to pass through quickly, thereby shortening the line measurement time as much as possible. 7th
As shown in the figure, the air layer 31a reaches the sensor 24,
The output signal is sent to a particle analysis circuit 35 and a liquid feeding means/control circuit 36. Liquid feeding means upon receiving the signal from the sensor 24]
to ensure that the sample liquid has an appropriate flow diameter in the flow cell section.
Apply proper pressure in the sample loop by pumping the syringe at normal speed. 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 to the recording medium is started, and at the same time, a signal is sent from the particle analysis circuit to the control circuit 36, and the sample pump 3 moves the syringe at high speed to move the sample loop 16.
The tube 18 and flow cell 19 are quickly cleaned by applying high pressure inside and rapidly feeding the cleaning sheath liquid.
The device stops after cleaning the waterway.

Various methods for performing particle analysis using histogram or cytogram processing based on the measurement data thus obtained are well known, and the calculations are performed in the particle analysis circuit 35.

[Another Embodiment Next, another embodiment of the present invention will be described using FIG. 8. In FIG. 8, the same reference numerals as in FIG. 1 represent the same members.

In FIG. 8, 50 is a compressor which is a source of compressed air, and an air tube connected to this compressor is branched into two branches. The branched air tubes are connected to a sheath liquid container 53 and a sample liquid container for airtightly storing the sheath liquid and sample liquid, respectively, via electric regulators 52 and 51 for pressure adjustment provided for the sheath liquid and sample liquid. It is connected to the liquid container 54.

The sheath tube 57 immersed in the sheath liquid in the sheath liquid container 53 is guided to the inflow portion of the flow cell 12 via the sheath liquid inflow control valve 58 . In addition, the sample liquid container 5
A translucent sample tube 55 immersed in the sample liquid in 4 is guided to the flow cell 12 via a sample liquid inflow control valve 59 . A waste liquid tube 60 is connected to the upper end of the flow cell 12, and the other end is connected to a waste liquid container 56. A sensor 61 for detecting air bubbles in the tube is attached near the sample liquid container 54 in the middle of the sample tube 55, and the output of this sensor 61 is connected to a control circuit 62 for liquid feeding means. The structure of the sensor 61 is similar to that shown in FIG. 2 described above. Liquid feeding means] The output signal of the control circuit 62 is connected to the regulators 51 and 52, the sample liquid inflow control valve 59, and the sheath liquid flow control valve 58, and controls the liquid feeding operation of the sample liquid and the sheath liquid.

In the above configuration, a predetermined pressurization value is applied to the sample liquid container 54 and the sheath liquid container 53 under the control of the regulators 51 and 52, and from the difference between the respective pressurization values, the flow cell 1
The sample flow diameter at 2 is determined. The sample in the sample liquid that sequentially flows through the test portion of the flow cell 12 is irradiated with a laser beam and measured, and the sample waste liquid that has been measured is discarded into the waste liquid container 56.

Here, if the sample liquid runs out while pressurizing the inside of the sample liquid container 54 and pushing out the sample liquid, or if the sample tube 61 is bent inside the container when the sample liquid container is attached, and the tip of the tube comes out above the liquid level. If it is closed, air is sent into the sample tube 55. Since the sample tube 55 is made of a translucent material, the refractive index inside the tube changes depending on the air sent, and the sensor 61
It is detected as a change in the output. If air in the sample tube is detected in this way and a sample liquid feeding failure is confirmed, the liquid feeding means] control circuit 62 controls the regulators 51 and 52, the sample liquid inflow control valve 59, and the sheath liquid flow control well 58. Then, the sample liquid feeding operation is stopped.

Note that the sensor 61 was installed near the sample liquid container 54 in the embodiment. However, in this case, if the sample liquid feeding operation is stopped at the same time as air in the tube is detected, the sample liquid present in the sample tube will be wasted at the time of stopping. Therefore, if the liquid feeding operation is continued for a certain period of time when air is detected by the sensor 61, and the operation is stopped after the sample liquid is fed into the flow cell 12, the sample liquid will not be wasted. Alternatively, if the sensor 61 is installed in the middle of the sample tube near the flow cell 12, the sample liquid will not be wasted.

The embodiments in which the present invention is applied to a flow cytometer have been described above, but the present invention is not limited to this, and can be applied to a particle counter, a Coulter measuring device that measures particulates from the electrical impedance of a sample, or an optical The present invention can be widely applied to devices that sequentially supply samples to a test area, such as a particle measuring device using acoustics.

[Effects of the Invention] According to the present invention, in the sample testing device, even if air is sent into the sample tube due to some accident during sample feeding, the feeding operation is stopped by detecting that air has been fed. Therefore, there is no adverse effect on the measurement.

[Brief explanation of the drawing]

1 and 3 to 7 are block diagrams of an embodiment of the present invention, FIG. 2 is a block diagram of a sensor for gas detection, and FIG. 8 is a block diagram of another embodiment of the present invention. This is a configuration diagram, and 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. A predetermined amount of sample liquid is supplied from a sample liquid container containing sample liquid to a test area by pressurization or suction, and the sample is tested in the test area. In the sample testing device, a detection means is provided in the middle of the sample supply route to detect gas in the sample supply route, and a means for pressurizing or suctioning when gas is detected by the detection means during supply of the specified amount. A sample inspection device comprising: control means for stopping. 2. The sample inspection device according to claim 1, wherein the detection means comprises a light source and a photodetector provided on both sides of the sample supply path. 3. The sample inspection device according to claim 1, wherein the sample is inspected by irradiating the supplied sample liquid with light and detecting an optical reaction thereof in the inspection section.