JPH04161836A - Micro quantitative dilution device - Google Patents

Abstract

PURPOSE:To determine and discharge a minute amount of sample liquid highly accurately by making an apparatus itself compact, and using a voltage expansion element and a diaphragm which is deflected by the expansion and the contraction of the element. CONSTITUTION:When a voltage is applied on a voltage expansion element 1 so as to elongate the element, a diaphragm part 2 is deflected in conformity with the movement of the element 1. The volume of a sample liquid well 13 which is formed in a central plate 5 of the main body of an apparatus for diluting a minute-amount of determined quantity is reduced. Thus, the sample liquid 3 is discharged into diluent liquid from the tip of a sample-liquid discharging nozzle 12 by the minute amount of the volume of the sample liquid well 13. The discharged sample liquid rides on the diluent liquid, passes through a valve 9 for mixed liquid and flows into a valve 15 for the mixed liquid. Therefore, a minute amount of the sample liquid corresponding to the deflection of the diaphragm part 2 can be accurately determined and discharged into the diluent liquid.

Description

[Detailed Description of the Invention] [Field of Industrial Application] The present invention relates to an apparatus for performing chemical analysis, biochemical analysis, or blood cell analysis, and in particular, the present invention relates to an apparatus for performing chemical analysis, biochemical analysis, or blood cell analysis. The present invention relates to a sample preparation device that performs large dilutions, and a sample microquantification device that quantifies a small amount of a sample with high precision.

[Conventional technology]

The quantitative dilution device for the sample liquid in the hematology analyzer includes:
For example, when measuring the number of blood cells per unit volume using the Coulter single method, such as the "Automatic Blood Analyzer" disclosed in Japanese Patent Publication No. 59-16667, the blood cells pass through minute holes, resulting in a high concentration. When measuring a blood sample, multiple blood cells may pass through the holes at the same time, or the holes may become clogged. To prevent this, a method is used in which whole blood is diluted several thousand to tens of thousands of times and the number of blood cells in this diluted solution is counted. The accuracy of this dilution is directly reflected in the accuracy of the measured value, so it is necessary to dilute with high precision.1 For example, in the above device, the dilution method uses a cut valve, which is a complicated quantitative sampling valve, to perform quantitative determination in two stages. It has been adopted.

[Problem to be solved by the invention]

In the conventional cut valve method described above, the amount of blood sample that can be quantified is at least several tens of microliters or more. Moreover, not only the cut valve device part but also the inside of the pipe leading blood thereto must be filled with blood. For this reason, the amount of blood used is several milliliters, which is quite a large amount of extra blood, compared to the several tens of microliters that is actually required for quantitative determination. Furthermore, since the amount of blood that is actually counted is about 10 nanoliters, it must be said that the several milliliters of blood required for dilution by the cut valve is a considerable waste. Furthermore, in this conventional technique, the diluent must also be quantitatively determined at the same time.

Further, in order to perform this large dilution, a large amount of diluent is required even if the dilution is performed in two stages.

The purpose of the present invention is to use micromachining technology to quantitatively quantify and dilute sample liquid in nanoliter units, thereby significantly reducing the amount of sample liquid required for measurement, and also reducing the amount of diluent required for dilution. It is an object of the present invention to provide a micro-quantitative dilution device that can significantly reduce the amount of dilution and can perform large dilutions with high precision without using complicated means such as two-stage dilution using a metering valve.

[Means to solve the problem]

In order to achieve the above object, the microquantitative dilution device of the present invention includes a sample liquid supply port, a sample liquid reservoir, a thin film part forming one surface of the sample liquid reservoir, and a voltage that is installed so as to be in contact with the thin film part. The voltage stretchable element is expanded and contracted by a voltage stretchable element drive device to cause deflection in the thin film part, thereby reducing the volume of the sample liquid reservoir and discharging a small amount of sample liquid. Quantitate the liquid.

Further, the micro-quantitative dilution device of the present invention has a diluent in the diluent flow path.
A sample liquid discharge nozzle for discharging a sample liquid is provided, and a dilution operation is performed by discharging the quantified sample liquid into a flowing diluent through the sample liquid discharge nozzle.

Furthermore, in the micro-quantitative dilution method of the present invention, the sample liquid is injected into the flowing diluent at the tip of the sample liquid discharge nozzle by adjusting the expansion and contraction operation of the voltage expansion and contraction element using a voltage expansion and contraction element drive device. Before and after discharging the sample liquid, the sample liquid is quantified with high precision by reproducing the same sample liquid shortage state.

Further, the blood cell counting device of the present invention includes the micro quantitative dilution device, performs micro quantitative dilution of blood using the micro quantitative dilution method, guides all of the diluted blood to the blood cell measurement section, and quantifies the blood. By counting all the blood cells in the blood, the number of blood cells per unit volume is measured.

Further, the microquantity metering device of the present invention comprises a sample liquid suction and discharge nozzle, a thin film portion forming one surface of the first working fluid reservoir, and one surface of the fifth working fluid reservoir, and a voltage expansion/contraction element installed so as to be in contact with the thin film portion. The voltage elastic element drive device expands and contracts the voltage elastic element to cause deflection in the thin film portion, thereby increasing or decreasing the volume of the sample liquid reservoir, and a minute amount of sample liquid is discharged from the sample liquid suction and discharge nozzle. .

[Effect]

The microquantitative dilution device of the present invention performs the following operations.

First, with the sample liquid valve open, the sample liquid passes through the sample liquid pipe, fills the sample liquid reservoir, and is further guided to the tip of the sample liquid discharge nozzle.

The diluent is flowing through the diluent pipe, and the sample liquid overflowing from the sample liquid discharge nozzle is discharged from the diluent outlet by the mixed liquid valve along with the flow of the diluent.

Next, the sample liquid valve is closed and the sample liquid is
Stand still with the sample liquid pipe, sample liquid reservoir, and sample liquid discharge nozzle filled to the tip. The mixed liquid valve is opened towards the mixed liquid pipe and the diluent flows into the mixed liquid pipe. In this state, a voltage is applied to the voltage expansion/contraction element to cause it to expand. When the voltage expansion/contraction element expands in response to a voltage, the diaphragm section bends in accordance with the movement, and the volume of the sample reservoir formed in the center plate of the main body of the microquantitative dilution device is reduced. This results in
The sample liquid is discharged into the sample liquid from the tip of the sample liquid discharge nozzle by an amount corresponding to the volume reduction of the sample liquid reservoir.

The discharged sample liquid.

The diluted liquid passes through the mixed liquid valve and flows into the mixed liquid pipe. Therefore, a very small amount of sample liquid corresponding to the deflection of the diaphragm portion is discharged into the diluent, and a dilution operation is performed. [Example] An example of the present invention will be described below with reference to the drawings.

FIG. 1 is an explanatory diagram of an embodiment of the microquantitative dilution device of the present invention. FIG. 2 is an explanatory diagram of the microquantitative dilution device of FIG. 1. FIG. 3 is a perspective view of the nozzle portion of the microquantitative dilution device of FIG. 1. FIG. 4 is an explanatory diagram of the sample liquid discharge state of the micro-quantitative dilution device of FIG. 1. 1 and 2, the main body of the microquantitative dilution device includes an upper plate 6, a center plate 5. It is constructed by joining three plates of the lower plate 7. These three plates are formed by micromachining, for example, from silicon single crystal plates, and have minute grooves and holes as shown in Figure 2.
Alternatively, a thin film is formed.

A diaphragm portion 2 is formed on the lower plate 7 of the main body of the micro-quantitative dilution device, and a voltage expansion/contraction element 1 is installed in contact with the diaphragm portion 2. A sample liquid reservoir 13 is formed in the center plate 5 of the main body of the microquantitative dilution device, and a sample liquid pipe 10. The sample liquid 3 is introduced through the sample liquid valve 8 .

A diluent pipe unit is connected to the main body of the micro-quantitative dilution device, and the diluted liquid 4 passes through the mixed liquid valve 9.
It is led to the diluted liquid outlet 14 or the mixed liquid pipe 15. Further, a sample liquid discharge nozzle 12 is formed in the center plate 5 of the main body of the microquantitative dilution device, and a third
As shown in the figure, the configuration is such that the sample liquid discharge position is in the center of the diluent liquid flow path.

The following operations are performed with the above configuration.

First, with the sample liquid valve 8 open, the sample liquid 3 passes through the sample liquid pipe 11, fills the sample liquid reservoir 13, and is further guided to the tip of the sample liquid discharge nozzle 12. The diluent 4 is flowing through the diluent pipe 11, and the sample liquid overflowing from the sample liquid discharge nozzle 12 rides on the flow of the diluent and passes through the mixed liquid valve 9.
As a result, the diluent is discharged from the diluent discharge port 14.

Next, the sample liquid valve 8 is closed, and the sample liquid 3
is the sample liquid pipe 11. Sample liquid reservoir 13. The sample liquid discharge nozzle 12 remains stationary with the sample liquid being filled to the tip. The mixed liquid valve 9 is opened toward the mixed liquid pipe 15, and the diluent 4 flows into the mixed liquid pipe 15.

In this state, a voltage is applied to the voltage expansion/contraction element 1 to cause it to expand. When the voltage expansion/contraction element 1 expands in response to voltage,
The diaphragm section 2 bends in accordance with the movement, and the volume of the sample liquid reservoir 13 formed in the center plate 5 of the microquantitative dilution device main body is reduced. Thereby, the sample liquid 3 is discharged from the tip of the sample liquid discharge nozzle 12 into the diluted liquid by a minute amount of the volume of the sample liquid reservoir 13 .

The discharged sample liquid passes through the mixed liquid valve 9 on the diluted liquid and flows into the mixed liquid pipe 15. Therefore, a minute amount of sample liquid corresponding to the deflection of the diaphragm portion 2 is discharged into the diluent.

Next, the timing of the operation of the voltage expansion and contraction element in the quantitative dilution operation will be explained in detail using FIG. 4. FIG. 4 is a diagram showing the sample liquid discharge state of the micro-quantitative dilution device of FIG. 1, in which the sample liquid reservoir 13 and the sample liquid discharge nozzle 12 are two-dimensionally shown.

In a, the sample liquid valve 8 is closed and the sample liquid 3
However, the sample liquid reservoir 13 and the sample liquid discharge nozzle 12 are filled to the tip. First, in this state, the voltage expansion/contraction element 1 is expanded by a minute amount to preliminarily eject the sample liquid. At this time, the amount of extension of the voltage expansion/contraction element is set so that a smaller amount of sample liquid is preliminarily ejected than the amount of sample liquid quantified. The sample liquid discharged at this time is carried away by the diluent flow 16.

Next, immediately following the operation in a, the voltage expansion/contraction element 1 is contracted by the amount expanded in the state in a (b).

At the tip of the sample liquid discharge nozzle 12, the flow of the sample liquid is cut off, and the diluted liquid enters the inside of the nozzle.

Next, the sample liquid cut off in the operation b is completely discharged from the diluted liquid outlet 14, and then the mixed liquid valve 9
is switched, and the movement of the voltage elastic element 1 is stopped until the diluent begins to flow toward the mixed liquid tube 15 (C).
At this time, diffusion occurs between the sample and the diluted liquid inside the sample liquid discharge nozzle 12, but this diffusion occurs only within the nozzle and does not spread to the flow path of the diluted liquid.

The amount of expansion of the voltage expansion/contraction element 1 at point a is adjusted in advance.

Next, the voltage expansion and contraction element 1 is expanded so that a fixed amount of the sample liquid is extruded. At this time, the amount of extension of the voltage expansion/contraction element 1 is adjusted so that a fixed amount of the sample liquid is discharged into the diluent together with the amount of the diluent sucked into the nozzle in the operation b.

Next, following the operation d, immediately contract the voltage expansion and contraction element 1 by the amount expanded in the state a (e),
Therefore, here, the state in which the sample liquid is cut off by the operation b is reproduced.

The amount of sample liquid discharged into the diluent is cut off by the operation b, and the remaining sample liquid is cut by the operation e,
Since the sample liquid is discharged into the diluent, highly accurate quantification is achieved by performing these two cutting operations in the same manner.

Performing a quantitative dilution operation using this micro-quantitative dilution device has the following advantages.

First, since the microquantitative dilution device is manufactured using micromachining technology, the device itself can be miniaturized, which allows the amount of sample liquid and dilution liquid required for dilution to be miniaturized. It is possible. In the case of blood tests, etc., it is desirable to collect as few blood samples as possible from a test subject, so a microquantitative dilution device is particularly useful when diluting blood.

Further, in the micro-quantitative dilution device described above, a voltage expansion and contraction element and a diaphragm that is deflected by the expansion and contraction operation of the voltage expansion and contraction element are used as means for discharging the sample liquid. This makes it possible to quantify a much smaller amount of sample liquid with higher precision than with conventional metering valves. Further, since a compressive force is directly applied to the sample liquid itself to perform the ejecting operation, the ejecting operation can be performed with good reactivity. Further, since the voltage expansion/contraction element can easily adjust the expansion/contraction amount by adjusting the voltage, the discharge amount of the sample liquid can be easily adjusted.

Next, another embodiment of the present invention will be described.

FIG. 5 is an explanatory diagram of another embodiment of the present invention.

In the present invention, the flow of the diluent is directed to the sample liquid discharge nozzle 12.
The shape of the hole in the center plate 5 of the microquantitative dilution device body is changed so that it is parallel to the diluent, so that the sample liquid flows as if being surrounded by the diluent. This results in
The sample liquid at the tip of the sample liquid discharge nozzle 12 is drained more accurately, and the accuracy of quantifying the sample liquid is improved. In the present invention, the diluted liquid outlet 14 and the mixed liquid tube 15 can be provided independently,
The flow of the diluent is controlled by the diluent valve 18 and the mixed liquid valve 9. This makes it possible to promptly discharge the preliminarily discharged sample liquid to the diluent discharge port.

Next, an example of a blood cell count measuring device using the micro-quantitative dilution device of the present invention will be described with reference to FIG.

FIG. 6 is an example of a blood cell count measuring device using the micro quantitative dilution device of the present invention. The main body of the micro-quantitative dilution device is fixed by a main body cover 19.

The diluent is sent to the main body of the micro-quantitative dilution device by the diluent supply device, and dilutes blood and feeds the blood. In order to perform this micro-quantitative dilution, a voltage elastic element driving device 26,
and an electromagnetic valve drive device 27 are installed. The mixed liquid made by this micro-quantitative dilution is transferred to the mixed liquid chamber 20.
sent to. Blood cells in the mixed liquid pass through the fine holes 22 for blood cell count measurement from the mother bell of the mixed liquid suction bag W123. At this time, a potential difference signal generated before and after the blood cell count measurement microhole 22 is detected by the blood cell count measurement electrode 21, processed by the blood cell count signal processing device 24, and passed through the blood cell count measurement microhole 22. Measure blood cell count. After all measurements are completed, sample tube 10. A cleaning liquid is flowed into the diluent tube 4, and the entire apparatus is cleaned.

An example of a blood cell count measuring device using the micro-quantitative dilution device of the present invention has the following advantages in addition to the advantages of the micro-quantitative dilution device shown in FIG.

First, since the amount of sample is very small and the dilution method is simple, there is no need to perform complicated procedures such as two-stage dilution using a conventional metering valve.

Furthermore, since the amount of the sample to be quantified is minute, it is possible to measure the number of blood cells contained in the entire amount of the quantified sample. This eliminates the need for quantitative determination of a diluent, which was necessary in the conventional blood cell count measurement method. Furthermore, since blood cells do not need to be evenly distributed in the liquid mixture to be measured, errors in blood cell count measurements due to uneven stirring that occur in conventional methods are eliminated.

Next, Figures 7 and 8 show an example of a microquantity metering device that uses a voltage stretchable element and a dial section that causes deflection.
This will be explained using figures. FIG. 7 is a principle diagram of a microquantity metering device that utilizes a voltage expansion/contraction element and a diaphragm portion that is deflected by the voltage expansion/contraction element. Moreover, FIG. 8 is an example of the micro-quantification device shown in FIG. 7.

The microquantity metering device according to the present invention has a voltage elastic element 1 and a diaphragm part 2 that is deflected by the voltage elastic element 1, and has a first
The device is substantially similar to the micrometer dilution device shown in the figure, but the micrometer metering device nozzle 31 protrudes into the atmosphere.

The following operations are performed with the above configuration.

First, the sample liquid pipe 11. and sample liquid reservoir 1
3 is filled with the hydraulic fluid 29. At this time, the voltage expansion and contraction element 1
is expanded to keep the diaphragm portion 2 in a deflected state.Next, the voltage expansion/contraction element 1 is slightly shortened to suck a small amount of divided air 30.

Next, the micro-volume quantitative device nozzle 31 is immersed in the sample liquid, the voltage elastic element 1 is shortened by a predetermined amount, and a predetermined amount of sample liquid is aspirated.Next, the micro-quantitative device nozzle 31 is installed at the sample liquid discharge location. , the voltage expansion/contraction element 1 is expanded by a predetermined amount, and a predetermined amount of sample liquid is discharged. The microquantity meter nozzle 31 has a shape that makes it easy to collect and discharge a small amount of sample liquid.

When there is a relatively large amount of sample liquid, with the sample liquid pipe 11 and sample liquid reservoir 13 completely filled with sample liquid, install the microquantity device nozzle 31 at the sample liquid discharge location, and set the voltage. The extensible element 1 may be extended by a predetermined amount and a predetermined amount of sample liquid may be discharged. next,
A cleaning liquid is flowed through the sample liquid pipe 1 to clean the entire apparatus.

Performing micro-quantification using this micro-quantification device has the following advantages.

First, since microquantity devices are manufactured using micromachining technology, the device itself can be made smaller.
This makes it possible to minimize the amount of sample liquid to be quantified. In addition, by making the nozzle 31 of the microquantity metering device finer, the error in liquid shortage at the tip of the nozzle can be made much smaller than in conventional metering devices. In addition, the microquantity meter uses a voltage expansion/contraction element and a diaphragm that is deflected by the expansion/contraction operation of the voltage expansion/contraction element as a means for discharging the sample liquid, so it is possible to perform a microdispensing operation with high quantitative accuracy.

Furthermore, since a compressive force is directly applied to the sample liquid itself to cause the discharge operation to be performed, it is possible to perform a suction and discharge operation with good reactivity. Further, since the voltage expansion/contraction element can easily adjust the amount of expansion/contraction by adjusting the voltage, it is possible to easily adjust the amount of suction and discharge of the sample liquid.

FIG. 8 is an example of the micro-quantification device shown in FIG. 7.

The main body 35 of this micro-quantification device is equipped with a dispensing switch 32, a dispensing amount setting switch 339, and a dispensing amount display section 34.The micro-dosing device uses electrical signals to perform quantitative determination using the above method. will be carried out. Therefore, it is possible to quantify a trace amount with high precision compared to the conventional quantitative method using manual operation.

〔Effect of the invention〕

According to the microquantitative dilution device of the present invention, by downsizing the device itself, the amount of sample liquid and dilution liquid required for dilution can be miniaturized.

In addition, by using a voltage expansion and contraction element and a diaphragm that is deflected by its expansion and contraction as a means for discharging the sample liquid, it is possible to meter out a minute amount of sample liquid with much higher accuracy than with a conventional metering valve. I can do it.

[Brief explanation of drawings]

FIG. 1 is an explanatory diagram of one embodiment of the micro-quantitative dilution device of the present invention, FIG. 2 is an explanatory diagram of the micro-quantitative dilution device of FIG. 1, and FIG. 3 is an explanatory diagram of the micro-quantitative dilution device of FIG. 1. FIG. 4 is an explanatory diagram of the sample liquid discharge state of the microquantitative dilution device of FIG. 1, FIG. 5 is an explanatory diagram of another embodiment of the present invention, and FIG. 1. A block diagram of a blood cell count measuring device using the micro quantitative dilution device of the present invention, FIG. The figure is a perspective view of the micro-quantification device of FIG. 7. DESCRIPTION OF SYMBOLS 1... Voltage expansion element, 2... Diaphragm part, 3...
... Sample liquid, 4... Diluent, 5... Center plate of the micro quantitative diluter main body, 6... Upper plate of the micro quantitative diluter main body, 7... Lower plate of the micro quantitative diluter main body, 8. ... Valve for sample liquid, 9... Valve for mixed liquid, 10... Tube for sample liquid, 11...
- Diluent tube, 12... Sample liquid discharge nozzle, 13... Sample liquid reservoir, 14... Diluted liquid outlet, 15... Mixed liquid pipe, 16... Flow of diluted liquid, 17...Diaphragm operation by voltage contraction element,
18... Valve for diluent, 19... Micro quantitative dilution device body cover, 20... Chamber for mixed liquid, 21...
・Electrode for blood cell count measurement, 22... Fine hole for blood cell count measurement,
23... Mixed liquid inhaler, 24... Signal processing device for blood cell count measurement, 25... Diluent supply device, 26... Voltage elastic element drive device, 27... Electromagnetic valve drive device,
28... General control device, 29... Hydraulic fluid, 30...
・Divided air, 31... Micro quantity metering device nozzle, 32...
・Dispensing switch, 3...Dispensing amount setting switch, 34・
... Dispensing amount display section, 35... Micro-quantity meter main body.

Claims (1)

[Scope of Claims] 1. A micro quantitative dilution device comprising a quantification part for quantifying a sample liquid and a dilution part for diluting it with a diluent, comprising: comprising a liquid reservoir, a thin film part forming one surface of the sample liquid reservoir, and a voltage expansion/contraction element installed in contact with the thin film part, and expanding/contracting the voltage expansion/contraction element by a voltage expansion/contraction element driving device; A microquantitative dilution device characterized in that the sample liquid is quantified by causing deflection in the thin film portion, thereby reducing the volume of the sample liquid reservoir, and discharging a small amount of the sample liquid. .