JPS6239697B2 - - Google Patents

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention provides a method for treating particles such as blood cells suspended in a liquid.
This is a particle detection device that can detect particles with high accuracy based on the difference in electrical impedance between liquid and particles.The previously aspirated sample is washed away after each measurement, and large and small particles such as red blood cells and platelets are mixed in. The present invention aims to provide a particle detection device that enables corrected counting of platelets in a mixed liquid.

[Conventional technology]

Conventionally, when counting particles such as blood cells, particles such as blood cells suspended in a liquid are detected by electrical or optical differences between the liquid and the particles, and then converted into an electrical signal and a pulse signal corresponding to the particle is detected. A method is used to count the number of

[Problem that the invention seeks to solve]

In conventional detectors, samples from the previous or earlier suction through the micropores remain, and those with particularly high specific gravity precipitate and fly up every time a measurement is made, causing a phenomenon of entrainment on the back side of the micropores. When you want to detect only platelets in a mixture of large and small particles, for example red blood cells and platelets, it becomes impossible to distinguish between pulses caused by larger red blood cell particles that enter the detection area and pulses from platelets. However, it had the disadvantage of giving errors to the measured values. Further, the number of pulses caused by the involvement of red blood cells can be determined from the number of red blood cells by a correction calculation, but it cannot be determined by a simple calculation due to the influence of the number of red blood cells in the previous or two previous samples.

As described above, it is quite difficult to classify and count particles in a suspension of particles in which two or more types of particles coexist. It is possible to use a method that prevents the so-called entrainment phenomenon by sucking all the particles passing through the micropores to the back side of the micropores, but this may make it impossible to quantify the sucked suspension. ,
Another drawback was that the structure became extremely complex. In addition, in the structure of a normal detector, it is generally directly connected to the quantitative part, and the liquid level is detected by using a mercury U-shaped tube or by providing a discontinuous part with the air layer. Although quantification is carried out, it has the drawback of contamination and corrosion caused by sample contamination, resulting in quantification errors and malfunctions.

The present invention has been made in order to eliminate the above-mentioned drawbacks, and aims to provide a particle detection device that can always perform measurements under the same conditions by washing out the previous sample sucked into the detector every time a measurement is performed. This is the purpose.

That is, in the prior art, for example, in order to provide a platelet count, it was necessary to correct the platelet count measurement result based on the current red blood cell count, the previous red blood cell count, and the previous red blood cell count. Furthermore, in order to avoid complicated correction formulas, it is necessary to avoid the influence of red blood cells, and the structure of the detector must be complicated. Furthermore, there is a problem in that the sample gets mixed into the quantitative part, causing stains and corrosion.

The present invention solves these problems.By washing and removing all previous samples, it is possible to correct the platelet count using only the current red blood cell count, which does not require a complicated detector. The platelet count can be determined using a simple correction formula without causing any problems or deviating the quantification of the aspirated suspension. Furthermore, by washing the metering section every time, the metering section is not contaminated and accurate quantification is achieved. The object of the present invention is to provide a particle detection device that can perform the following steps.

[Means for solving problems]

The particle detection device of the present invention will be described with reference to the drawings.
A detector pellet 7 having micropores 6 is provided in the part of the tube located in the sample liquid in the sample chamber,
A cleaning liquid supply pipe 1 is connected to the lower end of this pipe via a solenoid valve 8.
A cleaning liquid tank 13 is connected to the upper part of this cleaning liquid supply pipe via a liquid level adjustment control device 12. On the other hand, a liquid for quantifying the sample liquid is connected to the upper part of the pipe 4 passing through the sample chamber 1. Surface detection means 16,1
At the same time, an insulated relay part 18 that can freely communicate with the atmosphere and a negative pressure source is connected to the upper end of this tube, and electrodes are connected in the sample liquid 5 in the sample chamber 1 and in the tube 4 passing through the sample chamber. 24, 25 are arranged, and these electrodes, the liquid level detection means 16, 1
7 and the liquid level adjustment control device 12 are connected to a signal processing device 26 that classifies and counts particles of two or more sizes.

[Effect]

A cleaning liquid is passed through the tube 4 passing through the sample chamber for cleaning. At the same time, the sample in the sample chamber 1 is also discharged, and the next sample is introduced from the sample introduction port 2. The particle suspension is then drawn into the tube 4 through the fine holes 6 of the detector pellet 7. At this time, particles are detected based on the difference in electrical impedance between the liquid and the particles, and a pulse signal is generated. When the liquid level passes through the lower liquid level detection means 16, a counting start signal is generated, and when the liquid level passes through the upper liquid level detection means 17, a counting end signal is generated, and the number of particles during this period is calculated. counting and two liquid level detection means 16,
The number of particles per unit volume is measured by making the 17 sections a predetermined volume.

A tube 4 penetrating the sample chamber serves both as a quantitative tube and a cleaning tube. Since this tube 4 is washed by flowing a cleaning liquid before each measurement, the metering section between the liquid level detecting means 16 and the liquid level detecting means 17 is not contaminated and does not cause a metering error or malfunction. In this way, each washing washes out the previous specimen measurement sample from the tube 4, so counting is always started under constant conditions. For example, in the case of platelet counting, the red blood cells in the previous sample are This will no longer affect the current platelet count.

〔Example〕

Embodiments of the present invention will be described below based on the drawings. FIG. 1 shows an embodiment of the particle detection device of the present invention. Reference numeral 1 designates a sample chamber for containing a particle suspension, for example, a suspension of blood cells; the suspension of blood cells is introduced through the sample inlet 2, while the suspended suspension of blood cells that has already been measured is introduced through the sample outlet. It is discharged from 3.
It passes through this sample chamber 1 in the longitudinal direction and has an inner diameter of 3 mm.
A glass tube 4 of approximately 10 mm in diameter is provided, and a portion of the glass tube 4 located in the sample liquid 5 (particle suspension liquid) in the sample chamber 1 has a diameter of 70 to 100 μm, which is large enough to allow each blood cell to pass through. A detector pellet 7 having micropores 6 is provided. The lower end of the glass tube 4 is connected via a solenoid valve 8 to the lower end of a cleaning liquid supply pipe 10 provided vertically, and a liquid level adjustment control device 12 is connected to a liquid reservoir 11 at the upper part of this cleaning liquid supply pipe 10. A cleaning liquid tank 13 is connected. 14 is a liquid level detection device provided adjacent to the liquid reservoir 11;
Reference numeral 5 designates a solenoid valve provided in a pipe at the bottom of the cleaning liquid tank 13 for adjusting the liquid level. On the other hand, liquid level detection means 16 and 17 for quantifying the sample liquid are provided at regular intervals on the upper part of the glass tube 4 passing through the sample chamber 1, and the upper end of the glass tube 4 is connected to the atmosphere and a negative pressure source. A flexible insulating relay section 18 is connected. That is, 20 is a pipe communicating with the atmosphere, 21 is a pipe communicating with a negative pressure source, and 22 and 23 are electromagnetic valves. Further, an external electrode 24 and an internal electrode 25 are respectively arranged in the sample liquid 5 in the sample liquid chamber 1 and in the glass tube 4 passing through the sample chamber, and these electrodes 24, 25 and the liquid level detection means 16, 17 and the liquid level adjustment control device 12 are signal processing devices 2 that classify and count particles of two or more sizes.
6. Since the internal electrode 25 is on the hot side, the insulating relay part 18 is electrically disconnected to prevent leakage and noise from entering, and the structure is such that the upper tip of the glass tube 4 protrudes into the inside of the insulating relay part. None, the liquid in the glass tube flows out in the form of droplets. Therefore, the driving power source for the solenoid valve 8 is also of a floating type, and is used in a state where noise and leakage are prevented. On the other hand, the external electrode 24 is connected to the ground side and is immersed in the sample liquid 5 as described above.

In the particle detection device of the present invention configured as described above, the cleaning process is performed by the following operations. First, open the solenoid valves 8 and 23, and
When suction pressure is applied to the inside of the insulating relay part 18 through
The inside of the is cleaned. At the same time, the sample in the sample chamber 1 is also discharged, and the next sample is introduced from the sample introduction port 2. On the other hand, since the liquid level in the liquid reservoir 11 decreases, the cleaning liquid in the cleaning liquid tank 13 is absorbed by the solenoid valve 15.
The liquid is gradually introduced into the liquid reservoir 11 through the liquid reservoir 11 . When the specified cleaning is completed, the solenoid valve 23 is closed while the solenoid valve 8 remains open, and the solenoid valve 2 for opening to the atmosphere is closed.
2 is released. When the liquid level in the liquid reservoir 11 reaches the liquid level detection device 14, the liquid level adjustment control device 12 connected to the liquid level detection device 14 is activated to close the solenoid valve 15, and the supply of cleaning liquid is stopped. At the same time, the electromagnetic valves 8 and 22 are also closed, and liquid movement no longer occurs. The liquid level detection device 14 is provided slightly below the lower liquid level detection means 16, so the liquid level in this case is located directly below the lower liquid level detection means 16 in the glass tube 4. , is in a ready state.

The measurement process is performed by the following operations. First, only the solenoid valve 23 is opened to transmit suction pressure into the glass tube (the other solenoid valves are closed), and the particle suspension is sucked into the glass tube 4 through the fine holes 6 of the detector pellet 7. . At this time, particles are detected based on the difference in electrical impedance between the liquid and the particles, and a pulse signal is generated. The liquid level in the glass tube 4 gradually rises as the sample liquid is sucked through the fine holes 6 of the detector pellet 7, and when the liquid level passes through the lower liquid level detection means 16, a counting start signal is generated. When the liquid level passes through the upper liquid level detection means 17, a counting end signal is generated, the number of particles during this time is counted, and the two liquid level detection means 16, 17
By setting the section to a predetermined volume, the number of particles per unit volume is measured. When the liquid level reaches the upper liquid level detecting means 17 and the measurement is completed, the above-mentioned cleaning process is repeated again and measurement for another sample is started.

The above is a normal operation method for measuring relatively large single particles such as red blood cells or white blood cells, and since the inside of the glass tube 4, which corresponds to the inside of the detector, is cleaned every time, the liquid level detection means 16 of the liquid quantitative part is cleaned. ,1
7 malfunctions are prevented. Note that the solenoid valve 15 may be opened after all the liquid in the glass tube 4 has been drained first.

Furthermore, by configuring the signal processing device (circuit) 26 in the device of the present invention as shown in FIG. 2, a particle counting device can classify and count particles having two types of sizes such as platelets and red blood cells. can be provided. In FIG. 2, the output signal of the detection circuit 27 is converted to the level of red blood cells by the next gate circuit 30, so that the output signal of the detection circuit 27 is converted to the level of red blood cells by the threshold circuit 28, which has two threshold levels. Close the gate when the signal reaches
A signal is passed only to the red blood cell counting circuit 31 so as not to be counted as platelets, and on the other hand, only those at the platelet level are counted by the platelet counting circuit 32. Although a time lag occurs between the respective signals, by attaching a delay circuit 33 to the gate circuit 30 and a signal from a one-shot multivibrator to the gate signal 35 from the gate signal generation circuit 34, the red blood cell signal can be adjusted to match the platelet signal. Counting circuit 32
It doesn't reach. The number of red blood cells and the number of platelets are counted as described above, but since the number of platelets includes a signal due to the involvement of red blood cells, the next arithmetic circuit 36 calculates the number of red blood cells involved and a constant a ₀ to be described later. The true platelet count is calculated and displayed on the display circuit 37.

In this particle counting device, each detected signal is counted separately according to its height, and correction for large particles is made as follows: Number of small particles = Measured value of small particles - (a ₀ + a ₁ × number of large particles) Make it count. To explain in more detail,
In conventional particle detectors, it was found that the error caused by the pulse generation caused by the entrainment of large particles inside the detector on the counts of small particles is reproducible and is related to the number of large particles. . For example, when measuring a mixed solution of red blood cells and platelets, platelet count (PL) = (platelet count measurement result) - {a ₀ + a ₁ × (current red blood cell count RBC ₁ ) + a ₂ × (previous red blood cell count RBC ₂ ) +a ₃ × (red blood cell count RBC ₃ from the time before last) +...}. In other words, the results of the platelet measurement this time include pulses generated by the entrainment phenomenon inside the detector after the red blood cells that were mixed in this time are aspirated, pulses generated by the red blood cells in the sample that was previously aspirated, Since the pulses generated by the red blood cell entrainment phenomenon are counted as if they were platelet detection pulses, it is necessary to remove the pulses generated by these red blood cells. By determining the coefficients of the equation, it is possible to determine the platelet count that is close to the corrected true value.

When using the device of the present invention, in order to make the above equation simpler and more reliable, the process of setting a ₂ and subsequent values to zero, that is, the particle detection of the present invention shown in Figure 1 using the previous sample. All of it is cleaned and removed using a device, and the formula is as follows.

Platelet count (PL) = (Platelet count measurement result) - (a ₀ + a ₁ × RBC ₁ ) These a ₀ and a ₁ are greatly affected by the shape and structure of the detector, blood dilution factor, measurement time, etc. However, in the measurement example of this device, blood dilution ratio: 50,000 times Diameter of fine pores in detector pellet: 80 μ Counting time: 12 seconds Inner diameter of detector: 8 mm Measurement sample: 2.5 ml , a ₀ = 7.5 (10,000 pieces), a ₁ = 2.4/
100, RBC: (10,000 cells), good correlation with conventional platelet measurement methods (PRP method, etc.) was obtained.

〔Effect of the invention〕

Since the particle detection device of the present invention is configured as described above, the previous sample sucked into the detector is cleaned every time a measurement is performed, so measurements can always be performed under the same conditions, and the sample can be cleaned every time. By doing so,
The metering section does not get dirty and allows for accurate metering.
It also has the effect that the structure of the detector does not have to be so complicated. Furthermore, if a particle counting device is configured using the particle detection device of the present invention, the calculation formula will be simplified since it has a cleaning function as described above, and therefore particles of two or more sizes can be counted at once. This has the effect that classification and counting can be performed by measuring . For example, in the case of platelet counting, by washing and removing all of the previous sample, it is possible to correct the platelet count using only the current red blood cell count, and the platelet count can be determined using a simple correction formula.

[Brief explanation of the drawing]

FIG. 1 is a cross-sectional explanatory diagram showing one embodiment of the particle detection device of the present invention, and FIG. 2 is a systematic explanatory diagram showing an example of a signal processing device (circuit) in the particle detection device of the present invention. 1...Sample chamber, 2...Sample introduction port, 3...
...Sample outlet, 4...Glass tube, 5...Sample liquid,
6... Fine hole, 7... Detector pellet, 8... Solenoid valve, 10... Cleaning liquid supply pipe, 11... Liquid reservoir, 12... Liquid level adjustment control device, 13... Cleaning liquid tank, 14... ...Liquid level detection device, 15...Solenoid valve, 16, 17...Liquid level detection means, 18...Insulated relay section, 20...Air communication pipe, 21...Negative pressure source communication pipe, 22, 23... Solenoid valve, 24...external electrode, 25...internal electrode, 26...signal processing device,
27...detection circuit, 28...threshold circuit, 30...
Gate circuit, 31... Red blood cell counting circuit, 32...
Platelet counting circuit, 33...Delay circuit, 34...Gate signal generation circuit, 35...Gate signal, 36...
...Arithmetic circuit, 37...Display circuit.

Claims (1)

[Claims] 1. A tube is provided that penetrates the sample chamber, a detector pellet having micropores is provided in the part of this tube located in the sample liquid in the sample chamber, and a cleaning liquid supply tube is connected to the lower end of this tube via a solenoid valve. Connect the bottom end and
A cleaning liquid tank is connected to the upper part of this cleaning liquid supply pipe via a liquid level adjustment control device, and a liquid level detection means for quantifying the sample liquid is provided at the upper part of the pipe penetrating the sample chamber, and the upper end of this pipe is An insulated relay part that can be freely communicated with the atmosphere and a negative pressure source is connected to the insulating relay part, and electrodes are arranged in the sample liquid in the sample chamber and in a tube penetrating the sample chamber, and these electrodes, the liquid level detection means and A particle detection device characterized in that a liquid level adjustment control device is connected to a signal processing device that classifies and counts particles of two or more sizes.