JPH03202772A - Apparatus for washing reaction container - Google Patents

Abstract

PURPOSE:To accurately measure an antigen within a desired reaction time using a microplate by providing a nozzle lift means raising and lowering a suction nozzle so as to be capable of introducing the same into a reaction container. CONSTITUTION:A nozzle lift means 5 raising and lowering suction nozzles 4 so as to be capable of introducing the same in reaction containers 11 is provided to a washing apparatus. A washing solution is successively injected in the reaction containers 11 by washing nozzles 3 while microplates 1 are intermittently moved by the arrangement pitch of the reaction containers 11 at every predetermined time by a feed means 2. Further, the reaction solution or washing solution in the reaction containers 11 is sucked by the suction nozzles 4 in such a state that the suction nozzles 4 are introduced in the reaction containers 11 by the nozzle lift means 5. Thereafter, the suction nozzles 4 are drawn out of the reaction containers 11 by the nozzle lift means 5 to move the microplates 1 by the arrangement pitch of the reaction containers 11. Thereafter, by repeating this operation, the respective reaction containers 11 are washed. By this method, an antigen can be accurately measured within a desired reaction time.

Description

[Detailed Description of the Invention] <Industrial Application Field> The present invention relates to a cleaning device for reaction vessels, and more specifically, to automatically clean each reaction vessel of a microplate in which antigen-antibody reactions, etc. are carried out. The present invention relates to a reaction vessel cleaning device that can clean a reaction vessel.

<Prior art and problems to be solved by the invention> Conventionally, as one of the antigen measurement methods, EIA using antigen-antibody reaction has been used.
(enzyme immunoassay) is known. This E
An example of IA will be explained with reference to FIG. 7 showing a process diagram. First, a solid-phase antibody (or antigen) is injected into a reaction container, and an antigen (or antibody) to be measured is added thereto. After the antigen-antibody reaction is carried out, the inside of the reaction vessel is washed with a washing solution in order to remove the antigen (or antibody) that has not bound to the immobilized antibody. Next, an enzyme-labeled antibody is added to cause the antigen-antibody reaction to occur again, and then washing is performed with a washing solution to remove enzyme-labeled antibodies that have not bound to the antigen. Furthermore, after adding the substrate and injecting a reaction stop solution to stop the reaction, the amount of antigen can be determined by measuring the absorbance of each reaction container.

By the way, among the EIA mentioned above, in the microplate method using a microplate, reagents such as antibodies, antigens to be measured, labeled antibodies, etc. are automatically injected into each reaction container of the microplate, and then the relevant reagents are washed using a microwasher or the like. Cleaning is performed by automatically injecting a cleaning solution into the inside of the microplate.

However, in the microplate method described above, although a predetermined amount of reagent is sequentially injected into each reaction container, each reaction container is washed at the same time in microplate units. For this reason, the reaction time of the reagents differs between the reaction vessels of a single microplate, and there is a problem that only accurate measurements are possible under conditions where the reaction time is required to be constant. Kaatsu tko.

In order to solve this problem, a method has been adopted in which test tubes are used instead of the microplates mentioned above, and the reaction time of the reagent is measured for each test tube while washing is performed.
In this case, there is a problem that the number of steps required for the measurement is extremely large, and the equipment that automatically injects the reagent into the test tube and cleans the test tube becomes large.

In addition, as another antigen measurement method, instead of the enzyme-labeled antibody in the above EIA, the amount of antigen is measured by adding a fluorescent substance and determining the absorbance.The antigen is measured by adding FIA1 isotope and calibrating the γ-rays. RI to measure quantity
A and the like are known, but these measurement methods also have the same problems as the above-mentioned EIA. .

This invention was made in view of the above problems,
An object of the present invention is to provide a reaction container cleaning device that can accurately perform antigen measurement using a microplate in a desired reaction time.

Means for Solving the Problems> In order to achieve the above object, the reaction vessel cleaning apparatus of the present invention cleans a microplate in which a plurality of reaction vessels are arranged in series, and changes the arrangement pitch of the reaction vessels at predetermined intervals. a cleaning nozzle that sequentially injects cleaning waves into the reaction container of the microplate that is intermittently moved;
The apparatus is equipped with a suction nozzle that sequentially suctions the liquid in the reaction container, and a nozzle elevating means that raises and lowers at least the suction nozzle so that it can be introduced into the reaction container.

However, the reaction vessel cleaning device has a plurality of cleaning nozzles and suction nozzles arranged along the direction of movement of the microplate, and a control means for selectively injecting the cleaning liquid and suctioning the liquid. It may be further provided.

In the case of the reaction vessel cleaning apparatus having the above configuration, the cleaning liquid can be sequentially injected into each reaction vessel using the cleaning nozzle while the microplate is intermittently moved by the array pitch of the reaction vessels at predetermined intervals using the transport means. I can do it. Furthermore, with the suction nozzle introduced into the reaction container by the nozzle lifting means, the reaction liquid or washing liquid in the reaction container can be sucked by the suction nozzle, and then the suction nozzle is moved into the reaction container by the nozzle lifting means. The microplate can be moved by the array pitch of the reaction vessels. Thereafter, by repeating the above operations, each reaction container can be washed in sequence. Therefore, by intermittently moving the microplate in accordance with the difference in dispensing time of reagents between adjacent reaction vessels, it is possible to equalize the reaction time between each reaction vessel.

In particular, when a plurality of cleaning nozzles and suction nozzles are arranged along the moving direction of the microplate, and further comprising a control means for selectively injecting the cleaning liquid and suctioning the liquid, it is possible to The reaction time and the number of washings can be adjusted by regulating the injection of washing liquid into the reaction container and the suction of the washing liquid.

<Example> Hereinafter, embodiments will be described in detail with reference to the accompanying drawings showing examples.

FIG. 1 is a perspective view showing an embodiment of the reaction vessel cleaning apparatus of the present invention. The cleaning device includes a transport belt 2 as a transport means for intermittently moving the microplate 1, and a plurality of cleaning nozzles that are arranged along the transport direction of the microplate 1 and inject a cleaning liquid into the reaction container 11 of the microplate 1. 3, a suction nozzle 4 for sucking the reaction liquid and cleaning liquid in each reaction container 11, and a nozzle elevating means for introducing the cleaning nozzle 3 and suction nozzle 4 into each reaction container 11 and pulling them out from each reaction container 11. 5, and a control means 6 for selectively causing the cleaning nozzle 3 to inject the cleaning liquid and the suction nozzle 4 to suction the liquid.

As is well known, the microplate 1 is made up of a plurality of bottomed cylindrical reaction vessels 11 arranged in series, and the entire microplate 1 is integrally molded from transparent plastic.
(see figure).

The conveyor belt 2 is wound around a pulley (not shown) and is provided so as to pass directly below the cleaning nozzle 3 and the suction nozzle 4. This conveyor belt 2 is driven by a stepping motor so as to intermittently move the microplate 1 by the arrangement pitch A (see FIG. 2) of the reaction vessels 11. This conveyor belt 2 is controlled to be driven at time intervals equal to the time difference when dispensing a reagent such as an antigen from a negative reaction container 11 to a reaction container 11 adjacent to the reaction container 11. .

The cleaning nozzle 3 and the suction nozzle 4 are each 5 in the figure.
They are provided one by one, and form a pair for each reaction vessel 11. The cleaning nozzle 3 and the suction nozzle 4 are provided close to each other so that one set can be introduced into each reaction container 11 (see FIG. 3). However, the lower end of the cleaning nozzle 3 is set to be located above the lower end of the suction nozzle 4 by a predetermined height. Further, the cleaning nozzle 3 and the suction nozzle 4 each have a tube 31.
．． 41 is connected to the cleaning nozzle 3, and a cleaning liquid such as reaction stopping acid, physiological saline, pure water, etc. is supplied to the cleaning nozzle 3 from a cleaning liquid tank T1 through the tube 31 by a discharge pump P1. Ru. Further, the suction nozzle 4 is connected to a suction pump P2 via the tube 41, and a discharge port of the suction pump P2 is communicated with a recovery tank T2.

The nozzle elevating means 5 includes a clamp part 51 that clamps the cleaning nozzle 3 and the suction nozzle 4, an arm 52 connected to the clamp part 51, and a support part 53 that supports the arm 52 rotatably around a support shaft 53a. and an elevating section 54 that moves the support section 53 up and down.

The clamp portion 51 and the arm 52 are connected to the - group of nozzles 3.
．． 4 are provided independently. Further, the support shaft 53a of the support portion 53 is installed between a pair of support plates 53b which are arranged opposite to each other with each arm 52 in between, and each arm 52 is rotatably supported by the support shaft 53a. .

Further, the elevating section 54 can rotate a movable block 54b vertically movably supported by a pair of guide posts 54a, a screw member 54c screwed to the movable block 54b, and a screw member 54c in the normal and reverse directions. Rotationally driven motor 54
d. The arm 52 is set so that the horizontal state shown in FIG. 3 is the lower limit of its rotation range.

Furthermore, a pin 52a is vertically provided in the middle of each arm 52 to restrict the downward rotation of the arm 52 in cooperation with a stopper 62, which will be described later. The pins 52a are arranged in series along the direction in which the cleaning nozzles 3 and the suction nozzles 4 are arranged.

The control means 6 includes an electromagnetic on-off valve 61 (see FIG. 5) that controls the supply of cleaning liquid to the cleaning nozzle 3;
and a stopper 62 for regulating the descent of the suction nozzle 4. The electromagnetic on-off valve 61 is provided for each cleaning nozzle 3 so that the supply of cleaning liquid can be individually controlled. Further, the stopper 62 is a drum-shaped member that is rotatably supported around a shaft 62b arranged along the arrangement direction of the cleaning nozzle 3 and the suction nozzle 4, and has a part on its outer periphery that is perpendicular to the arm 52. Pin holes 62a are arranged into which the provided pins 52a can be introduced.

The pin holes 62a are arranged along each of a plurality of generatrix lines L1 to L8 set at equal intervals in the circumferential direction of the stopper 62, and their formation pattern is
-L8 (see Fig. 6). For example, on the generatrix Ll, they are formed for each line segment X1 to X5 in the circumferential direction corresponding to the axial center position of the pin 52a, and on the generatrix L2, is formed only on the circumferential line segment X5, and on the generatrix L3, it is formed only on the circumferential line segments X5 and X4. Further, the stopper 62 is rotated by a stepping motor (not shown) so that one of the patterns of the bottle holes 62a is at the top. Rotation control of the stopper 62 by this stepping motor is performed by manually inputting a desired pattern of pin holes 62a to a microcomputer. Here, the desired pin hole 6
With pattern 2a selected, the movable block 54
When b is lowered, downward movement of each arm 52 is permitted or restricted in accordance with the pattern of the pin holes 62a. That is, among the plurality of arms 52, when the movable block 54b is lowered, the pin 52a is inserted into the hole 62a.
Only those introduced into the microplate 1 can move downward together with the movable block 54b, and in turn, the cleaning nozzle 3 and the suction nozzle 4 supported by the arm 52 can be introduced into the reaction container 11 of the microplate 1. Yes (see Figure 3). Further, in the arm 52 in which the pin 52a is not introduced into the pin hole 62a, the pin 52a is inserted into the stopper 62a.
As the elevating part 54 descends, the arm 52 is relatively rotated clockwise as shown in FIG. 4, and the cleaning nozzle 3 and suction nozzle 4 supported by the arm 52 are introduction into the reaction vessel 11 is regulated.

At this time, the corresponding electromagnetic on-off valve 61 is maintained in a closed state so that the cleaning liquid is not discharged from the cleaning nozzle 3 whose introduction into the reaction vessel 11 is restricted. The opening and closing of the electromagnetic on-off valve 61 is controlled by the microcomputer according to the selected formation pattern of the pins 52a.

The operation of the reaction vessel cleaning device having the above configuration is as follows. First, the microplate 1 is transported by the conveyor belt 2.
When the reaction container 11 located at the downstream end in the transport direction is moved directly below the pair of nozzles 3.4 located at the upstream end in the transport direction, the screw member 54c is rotated. , the movable block 54b descends. As for the arm 52 whose pin 52a is introduced into the pin hole 62a of the stopper 62, its cleaning nozzle 3 and suction nozzle 4 are introduced into the reaction container 11 of the microplate 1 at a constant speed. During this introduction, the reaction liquid in the reaction container 11 is sequentially sucked by the suction nozzle 4 and collected into the collection tank T2. When suction of the reaction liquid is completed, screw member 5
4c is reversed, and the cleaning nozzle 3 and suction nozzle 4 are raised. Then, when the cleaning nozzle 3 rises to near the mouth edge of the reaction vessel 11, the rising of each nozzle 3, 4 is stopped, and the cleaning nozzle 3 is placed inside the reaction vessel 11.
Cleaning liquid is injected through the After that, each nozzle 3 again
, 4 are lowered, and the cleaning liquid in the reaction container 11 is sucked by the suction nozzle 4 and collected into the collection tank T2. Thereafter, after the above operation is repeated for a predetermined period of time, each nozzle 3.4 is pulled out from the reaction vessel 11.

Next, the microplate 1 is intermittently moved by the formation pitch A of the reaction vessels 11 by the conveyor belt 2, and the reaction vessel 1 is positioned at the downstream end of the microplate 1 in the conveyance direction.
1 is transported directly below a pair of nozzles 3 and 4 located second from the upstream end in the transport direction, and at the same time, the reaction container 11 adjacent to the reaction container 11 is transported to the upstream end in the transport direction. The nozzle 3.4 is moved directly below the nozzle 3.4. The nozzle 3.4 is then introduced into the reaction vessel 11, and the reaction vessel 11 is cleaned in the same manner as described above. Thereafter, the above operation is repeated for each reaction vessel 11, and cleaning of the reaction vessel 11 is completed.

In this way, according to the cleaning device, the reaction vessels 11 can be sequentially cleaned in accordance with the difference in dispensing time between the reaction vessels 11 of the microplate 1, so that the reaction between the reaction vessels 11 can be reduced. You can even out the time.

On the other hand, for the arm 52 in which the pin 5.2g is not introduced into the pin hole 62a of the stopper 62, the nozzle 3.4 supported by the arm 52 is prevented from being introduced into the reaction vessel 11, and the electromagnetic on-off valve 61 The injection of the cleaning liquid is stopped, and the cleaning inside the reaction vessel 11 is regulated. Therefore, by appropriately selecting the formation pattern of the pin holes 62a of the stopper 62, it is possible to adjust the reaction time and the number of washings for a desired reaction container 11.

In the above embodiment, the - group of nozzles 3°4 is
Although they are integrally raised and lowered by one arm 52, they may be arranged alternately along the transport direction of the microplate 1 and individually raised and lowered by a single arm 52.

Further, the number of nozzles 3.4 can be increased or decreased as appropriate. For example, if the purpose is to make the reaction time uniform among the reaction vessels 11, the number of nozzles 3.4 can be increased or decreased as appropriate. It suffices if at least one set is provided.

In addition, the reaction container cleaning device of the present invention is not limited to the above embodiments, and for example, as a conveying means,
Instead of the conveyor belt 2, one or more pairs of conveyor rollers that are driven intermittently while sandwiching the sides of the microplate 1 can be used, and the mouth edge of the reaction vessel 11 can be moved without raising or lowering the cleaning nozzle 3. Numerous design changes can be made, such as fixing it slightly upward.

Furthermore, the reaction vessel cleaning apparatus of the present invention can be used in various cleaning processes in FIASRIA in addition to EIA.

<Effects of the Invention> As described above, according to the reaction vessel cleaning device of the present invention, the reaction vessels can be sequentially cleaned according to the time difference when dispensing reagents to each reaction vessel of a microplate. Therefore, the reaction time between the reaction containers can be made uniform, and the amount of antigen, etc. can be measured accurately.

Moreover, since a microplate is used, measurements can be carried out efficiently and the apparatus can be made smaller.

In particular, when a plurality of cleaning nozzles and suction nozzles are arranged along the moving direction of the microplate, and a control means for selectively injecting the cleaning liquid and suctioning the liquid is provided, each As a result of being able to individually control the reaction time of the reagent and the number of washings of the reaction container, a unique effect is achieved in that the amount of antigen, etc. can be measured easily and efficiently under various conditions.

[Brief explanation of drawings]

FIG. 1 is a perspective view showing an embodiment of the reaction container cleaning device of the present invention, FIG. 2 is a perspective view of a microplate, FIG. 3 is a side view showing a state in which a nozzle is introduced into the reaction container, and FIG. Figure 4 is a side view showing a state in which the introduction of the nozzle into the reaction vessel is restricted, Figure 5 is a schematic diagram showing the piping system, Figure 6 is a developed view showing the pin hole formation pattern of the stopper, Figure 7 is a process diagram of EIA. 4... Suction nozzle, 5... Nozzle lifting means, 6...
- Control means.

Claims (1)

[Claims] 1. A transport means for intermittently moving a microplate in which a plurality of reaction vessels are arranged in series by the arrangement pitch of the reaction vessels at predetermined time intervals; Cleaning of a reaction vessel characterized by comprising a cleaning nozzle for injecting a cleaning liquid, a suction nozzle for sequentially suctioning the liquid in the reaction vessel, and a nozzle elevating means for raising and lowering at least the suction nozzle so as to be introduced into the reaction vessel. Device. 2. The microplate according to claim 1, further comprising a plurality of cleaning nozzles and suction nozzles arranged along the moving direction of the microplate, and a control means for selectively injecting the cleaning liquid and suctioning the liquid. Reaction vessel cleaning equipment.