JPH02296131A - Particle counting apparatus - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed Description of the Invention] [Field of Industrial Application] The present invention relates to a particle counting device that measures the number of particles and performs particle analysis using light, electric fields, magnetic fields, etc. by flowing a liquid containing particles into a capillary channel. Regarding.

[Conventional technology]

One of the conventional particle counting devices is a blood cell classification device.
There are some that are listed in the publication. This device consists of a red blood cell measurement system that collects a fixed amount of whole blood, then sends a predetermined sample solution that has been diluted quantitatively to a flow cell to measure the number of red blood cells in the fixed solution, and a predetermined sample solution that has been diluted in place at a dilution rate different from the above. The system consists of a white blood cell measurement system that measures the number of white blood cells and their subpopulations in a given solution using a flow cell by mixing reagents such as hemolytic agents or staining agents with the sample solution. In conventional apparatuses, the so-called specimen pretreatment process described above is performed while a sample liquid is sent through a pretreatment system connected by a tube. Therefore, in order to divide one test sample into a red blood cell measurement system and a white blood cell measurement system and efficiently perform pretreatment for each, a quantitative sampling valve and numerous electromagnetic valves and pumps are connected by complicated tubing.

[Problem to be solved by the invention]

The above conventional technology is used to transport samples and reagents in a blood cell sorting device to a flow cell through valves and tubes, but the samples and reagents accumulate on the inner walls of the tubes and at the joints with the valves. There was a problem in that measurement results were adversely affected by clogging and mutual contamination.

Another problem is that the pretreatment system, in which the flow of samples and reagents is fixed by tubing, lacks versatility for other adjustment processes.

The purpose of the present invention is to solve the above-mentioned problems and provide a versatile particle counting device that can eliminate clogging and cross-contamination in tubes and valves, and can perform various adjustment processes. There is a particular thing.

[Means to solve the problem]

In order to achieve the above object, the particle counting device of the present invention is a particle counting device that performs particle counting 411 using a sheath flow cell, and a particle counting device that temporarily stores sample samples and performs adjustments such as staining, hemolysis, and dilution. A reaction vessel, a sample sample alone or a sample sample and reagents are aspirated and discharged into the reaction vessel, and then the conditioned sample from this reaction vessel is aspirated and is directly connected to the sample nozzle on the outer surface of the sheath flow cell. The first pipettor is connected to the sample supply port and discharges the adjusted sample.

Further, in place of the first pipettor, a second pipettor is provided which performs only the operation of sucking only the sample sample or the sample sample and the reagent and discharging them into the reaction container.

Also, instead of the first pipettor or the second pipettor,
It is equipped with a third pipettor that sucks in the reagent and discharges it into the reaction container.

Furthermore, it is provided with only a plurality of the necessary parts of the above-mentioned particle counting device.

[Effect]

The first pipettor of the particle counting device aspirates the sample sample and reagent in this order or the reverse order and then transports them to the reaction vessel and dispenses them, or if the reagent is used as a working fluid for the dispensing suction. Only the sample sample is aspirated, and the reagent is discharged following the sample sample during dispensing.The first reaction container temporarily accommodates these and makes adjustments such as staining, hemolysis, dilution, etc., and then the first pipettor is used to carry out this reaction. Aspirate the adjusted sample from the container, transport it to the sheath flow cell, connect it to the sample supply port on its outer surface that communicates with the internal sample nozzle, and discharge the aspirated adjusted sample to the sheath flow cell. Inside, the sheath liquid is supplied in time for this discharge to form a sheath flow, and the particle meter 8I'l is performed here.

In addition, the second pipettor of the particle measuring device aspirates the sample sample or the sample sample and reagent, moves them to the reaction container, and discharges them, and the first pipettor aspirates the prepared sample from the reaction container and inserts it into the sheath. This is carried to the flow cell and supplied as a sheath flow sample liquid.

In addition, the third pipettor of the Will particle analyzer aspirates and dispenses the reagent, the second pipettor aspirates and dispenses the sample sample, and the first pipettor supplies the adjusted sample to the sheath flow cell. do.

Each of the above particle meters? Due to the action of the l11 device, sample samples, reagents, and adjustment samples only pass through a pipettor on the way to the sheath flow cell, which eliminates clogging and cross-contamination in tubes and valves that were seen in conventional technology. Furthermore, since the process of sample adjustment using a vivette can be easily changed, it is possible to perform various adjustment processes compared to the conventional method.

Furthermore, the above particle meter? ! ! In an I-device equipped with a plurality of necessary parts, such as a reaction vessel, the same sample is subjected to different adjustment processes according to the operating procedures of each pipettor, and then supplied to the sheath flow cell.

Due to the above-mentioned effect, it is possible to completely eliminate cross-contamination between each reagent.

〔Example〕

Embodiments of the present invention will be described below with reference to FIGS. 1 to 5.

FIG. 1 is a perspective view 1+4 showing a first embodiment of a particle counting device according to the present invention. In FIG. 1, a tapered sample supply port 10 communicating with a sample liquid nozzle inside the flow cell is provided in the second part of the sheath flow cell 9. A cleaning tank 8 for cleaning the first pipettor 1 is provided adjacent to the sheath flow cell 9, and then the sample is temporarily stored for staining, hemolysis, dilution, etc. 111! A reaction volume fI3, a reagent container 4, and a sample volume H#5 are provided in this order along a certain circumference. The first pipettor 1, which transports the sample, is installed so as to move j2 times along the circumference. A lens 13 for condensing laser light, a laser light source 12, and a cage are provided in this order in front of the sheath flow cell 9, and a lens 14 for concentrating scattered light and fluorescence and a light detector 19 are provided on the opposite side of the sheath flow cell 9. They are provided in this order.

The above configuration works as follows. The pipettor 1 @1 first descends into the reagent container 4 and aspirates a predetermined amount of the reagent 7. Then, as shown by the arrow 20, it moves into the sample container 5 and aspirates a predetermined amount of the sample 6. Thereafter, as shown by arrow 21, it descends into reaction vessel 3 and discharges sample sample 6 and reagent 7. Also, in some cases fjs
The above-mentioned suction and ejection may be repeated using the pipettor L to promote stirring and staining.Next, after a certain period of time has elapsed, this iW prepared sample 11 is aspirated and the arrow 22
As shown, move to the sample supply port 10 and connect the O-ring 2 at the top of the first pipettor 1 and the sample supply port 10.
connects with the inner surface of By supplying the sheath liquid while maintaining this state and slowly discharging the prepared sample 11, a sheath flow 23 can be formed within the sheath flow cell 9.

When the particle number measurement and particle analysis using the laser light source 12, laser beam focusing lens 13, scattered light/fluorescence focusing lens 14, and optical detector 19 are completed, the first pipettor 1 returns to the reaction vessel. Return to step 3 and thoroughly discharge clean water to clean the inside of the container.

Thereafter, all of these cleaning liquids are sucked up, moved to the cleaning tank 8, and discharged, thereby cleaning the first pipettor 1 itself.

FIG. 2 is a partial sectional view for explaining a part of the first pipettor 1 in relation to FIG. 1. In the embodiment shown in FIG. 1, a reagent container 4 is provided, but as shown in FIG. It is possible to supply a predetermined amount of the sample sample 6 and reagent 7 in the same manner as in the above embodiment.

FIG. 3 is a top view showing a second embodiment of the particle counting device according to the present invention. In FIG. 3, this embodiment shows the first
A second pipettor 24 is provided to carry out the suction and discharge operations of the reagent 7 and sample sample 6 of the first pipettor 1 in the first embodiment shown in the figure. A sample supply port 10 of a sheath flow cell 9, a reaction vessel 3, and a cleaning tank 8 are installed in this order along a first circumference, and the first pipettor 1 can rotate on this first circumference. It is arranged like this. Further, the reaction container 3, the cleaning tank 8, the reagent container 4, and the sample container 5 are arranged along the second circumference. That is, the first circumference and the second circumference intersect at the reaction vessel 3 and the washing tank 8, and the second pipetter 24 is arranged so as to be able to rotate along this second circumference. Ru. Other details are the same as in FIG.

The above configuration works as follows. First, use the second pipettor 2.
4 moves to the position of the reagent container 4 as shown by the arrow 30 and descends to aspirate the reagent 7, and then moves to the sample container 5 as shown by the arrow 25 and descends to aspirate the sample sample 6. These are then moved as indicated by arrow 26 and discharged into the reaction vessel 3 in a predetermined amount. In some cases, the above suction and discharge may be repeated to promote the reaction. Thereafter, as shown by arrow 327, the first pipettor 1 moves to the washing tank 8 and performs washing. At the same time, the first pipettor 1 located above the sample supply port 10 moves to the reaction container 3 as shown by arrow 29 and descends. The adjusted sample 11 is aspirated, connected to the sheath flow cell 9 in the same manner as in FIG. 1, and sample discharge for measurement is started.

When the measurement of the number of particles using the laser light source 12, lenses 13, 14, and optical detector 19 is completed, the reaction vessel 3 is cleaned in the same manner as in FIG. 1, as shown by the arrow 29. After that, when the cleaning is finished, the first pipettor 1 moves to the cleaning tank 8 for cleaning itself as shown by the arrow 31, but at this time the second pipettor 24 is already in operation for the next sample 6. In the washing tank 8, the first pipettor 1 and the second pipettor 24 perform zero or more operations without colliding once or multiple times to analyze one particle.

FIG. 4 is a top view showing a third embodiment of the particle counting device according to the present invention. In FIG. 4, this embodiment shows the third
A third pipettor 34 is provided to carry out the suction and discharge operations of the reagent 7 of the second pipettor 24 in the second embodiment shown in the figure. The cleaning tank 8 of the first pipettor 1 and the reaction vessel 3 are arranged in this order along the first circumference, and the first pipettor 1 is connected to the sample supply port 10 of the sheath flow cell 9.
It is arranged so that it can rotate on the circumference of Also, the first
The reagent container 4, the reaction container 3, and the second cleaning tank 35 are arranged in this order along a second circumference that intersects the circumference of the reagent container 3 at the reaction container 3. It is arranged so that it can rotate on the circumference of 2. Further, a third circumference intersects the second circumference at the reaction vessel 3 and the second cleaning tank 35.
The reaction container 3, the second washing tank 35, and the sample sample container 5 are arranged along the circumference of the third circle, and the second pipetter 24 is arranged so as to be able to rotate on the third circumference.

The above configuration works as follows. First, use the second pipettor 2.
4 descends into the sample container 5, sucks up the sample 6, and discharges it into the reaction container 3. Thereafter, the process moves to second cleaning M35, where the second pipettor 24 itself is cleaned, and then moved to the sample sample container 5. At the same time, the third pipettor 34 sucks the reagent 7 from the reagent container 4 and discharges it into the reaction container 3. Thereafter, it moves to the second cleaning tank 35 to wash the third pipettor 34 itself, and then moves to the reagent container 4. In some cases, the reaction may be promoted by repeating the suction and discharge operations of the sample 6 and reagent 7 into the reaction container 3. During this time, the first pipettor 1 moves to the reaction vessel 3, aspirates the prepared sample 11, carries it to the sheath flow cell 9, and d(
start setting.

When this is completed, the pipette moves to the reaction container 3 again to wash the container, and then moves to the washing tank 8 to wash the first pipettor 1 itself. The above operations are repeated one or more times to perform particle analysis.

FIG. 5 is a top view showing a fourth embodiment of the particle counting device according to the present invention. In FIG. 5, this embodiment has five reaction vessels 40, 41, 42, 43, 44 arranged in this order on the circumference, whereas the above embodiment was equipped with one reaction vessel 3. It is equipped with a rotary table 45.

For the first pipettor 1, there is a sample supply port 10 of the sheath flow cell 9, a washing tank 8, and a reaction container 40 in the figure, and for the twentieth pipettor 34, there is a reagent container 42,
There is a third cleaning tank 38 and a reaction vessel 42, and a second
The sample sample container 5 and the second pipettor 324 are
There are a cleaning tank 35 and a reaction container 43. Furthermore, there is a stirrer lock 36 for stirring the prepared sample 11 in the reaction vessel 41.

A fourth cleaning tank 4 is provided for cleaning it. Further, a cleaning mechanism 39 for sucking the remaining adjusted sample sample 11 in the reaction container 44 and cleaning the inside of the reaction container 449 is connected to the reaction container 449.
It is located at

The above configuration works as follows. First, use the second pipettor 2.
4 sucks the sample 6 in the sample sample container 6 and discharges it into the reaction container 43. Then use the second pipettor 2
4 moves to the second washing tank 35 to wash itself, and when the washing is completed, moves above the sample sample container 5 and waits for the next sample 6. During this time, the reaction rotary table 45 rotates by 72″ instead of clockwise, and the reaction vessel 4
3 to the position of the reaction vessel 42 in the figure. At this position, the third pipettor 34 aspirates the reagent 4 in the reagent container 4 and adds it to the sample 6 in the reaction container 43. After washing the third pipettor 34 itself in the third washing tank 38, 4 and waits for the next reaction container 44 to come around. During this time, the reaction rotary table 45 rotates 72 clockwise.
'' to move the reaction container 43 to the position of the reaction container 41 in the figure. At this position, the stirring mechanism 36 stirs the adjusted sample 11 in the reaction container 43, and when the stirring is completed, it moves to the fourth cleaning tank 37. After moving and cleaning the stirring mechanism 36 itself, the reaction rotary table 4 waits for the next reaction container 44.
5 rotates 721 times clockwise to move the reaction container 43 to the position of the reaction container 40 in the figure. At this position, the first pipettor 1 aspirates the adjusted sample 11 in the reaction container 43 and supplies it to the sheath flow cell 9. Here, laser light source 1
2, lenses 13 and 14, and an optical datater 19 to measure the number of particles. After that, the first pipettor 1 washes itself in the washing tank 8 and waits for the next reaction vessel 44. During this time, the reaction rotary table 45 rotates clockwise by 7
2" rotation to move the reaction container 43 to the position of the reaction container 44 in the figure.

At this position, the cleaning mechanism 39 cleans the reaction vessel 43, and then the reaction vessel 43 is moved to the position of the reaction vessel 43 in the figure. The above operation is carried out in each reaction vessel 40, 41, 42,
By continuously performing 43 and 44, it is possible to pre-process many specimens, measure the number of particles, and perform particle analysis in a short period of time.

〔Effect of the invention〕

According to the present invention, sample samples, reagents, and adjustment samples from a single-particle measurement device only pass through a pipettor on the way to the sheath flow cell, eliminating clogging and cross-contamination in tubes, valves, etc. that were seen in conventional technology. In addition, since the process of sample adjustment using a pipettor can be easily changed, it is possible to perform various adjustment processes compared to the conventional method.

[Brief explanation of drawings]

FIG. 1 is a block diagram showing a first embodiment of a particle counting device according to the present invention, FIG. 2 is a partial sectional view of a first pipettor related to FIG. 1, and FIG. FIG. 4 is a block diagram showing a third embodiment of the present invention, and FIG. 5 is a block diagram showing a fourth embodiment of the present invention. DESCRIPTION OF SYMBOLS 1... First pipettor, 3... Reaction container, 4...
Reagent container, 5... Sample sample container, 9.° G-S flowal 10... Sample supply port, 12... Laser light source, 19... Optical detector, 24... Second pipetter, 34 ...Third pipettor, 45...Reaction table. No. 1 Kanto Sugar No. Σ

Claims (1)

[Claims] 1. In a particle counting device that measures particles using a sheath flow cell, sample samples are temporarily stored, stained,
A reaction container for making adjustments such as hemolysis and dilution, a sample sample alone or a sample sample and a reagent are aspirated and discharged into the reaction container, and then the adjusted sample is aspirated from the reaction container and a sheath is inserted. 1. A particle counting device comprising: a first pipette connected to a sample supply port on the outer surface of the flow cell that communicates directly with a sample nozzle, and for discharging an adjusted sample. 2. In place of the first pipette, a second pipette is provided which performs only the operation of suctioning only the sample sample or the sample sample and the reagent and discharging them into the reaction container. particle counting device. 3. Instead of the first pipette or second pipette,
3. The particle counting device according to claim 1, further comprising a third pipette that performs only the operation of sucking the reagent and discharging it into the reaction container. 4. A particle counting device comprising a plurality of the necessary parts of the particle counting device according to claim 1, claim 2, or claim 3.