JPH06194315A - Continuous titration emission measurement system - Google Patents

Abstract

PURPOSE:To shorten the work time by moving a nozzle for dispensing an emission source substance generating reagent in X-Y direction along the surface of a multi-well plate. CONSTITUTION:A multi-well plate 5 arranging a large number of titration containers in matrix is mounted on an X-Y drive table 6. A nozzle 1 for dispensing emission substance generating reagent sequentially to respective titration containers is provided and driven in X-Y direction on a plane parallel with the plane of the plate 5 to be driven. To a titration container, for which a predetermined incubation time has ended, with an emission reagent is then injected by an emission reagent injection nozzle 2 through a syringe unit 16. Consequently, a sample in the titration container begins to emit light. The light is taken out by a lens unit 4 arranged closely to the nozzle 2 integrally therewith and introduced through an optical fiber 17 to a light receiving element 4'. An output signal from the light receiving element 4' is then converted into a numerical electric signal by an electronic circuit 18 and processed by means of a computer 19.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a continuous titration apparatus, and in particular, to a multiwell plate in which a large number of titration vessels are arranged vertically and horizontally, a reagent is successively and continuously injected to emit light from a sample in the titration vessel. The present invention relates to a continuous titrator which is convenient for use in, for example, basic research on bioluminescence / chemiluminescence, research on the activity of living cells, clinical tests (immunoassay), and measurement of the number of bacteria in food.

[0002]

2. Description of the Related Art A multi-well plate in which a large number of titration vessels are arrayed vertically and horizontally is put in each of the titration vessels, and a sample having slightly different properties is put into the titration vessel to measure and analyze the luminescence of each titration vessel. Have existed in the past, they are dispensed to each titration container containing a sample, the luminescent source substance-producing reagent is dispensed, the sample and the luminescent source substance-producing reagent are incubated, the luminescent reagent is injected, and the luminescent measurement and Five operations of injecting a stop reagent are performed in this order for a predetermined time. However, the number of titration containers is generally 24, 48, or 96, and even if it is automated, the total working time is enormous.

[0003]

The continuous titration apparatus of this kind usually has a self-supporting work. However, since the total working time is lengthened, the number of samples that can be measured per day is increased. In addition, the sample may change over time, resulting in an error in the data. Therefore, the present invention is intended to reduce the working time.

[0004]

According to the present invention, the most time-consuming operation among the above-mentioned five operations must maintain the constant temperature of the container after the injection of the luminescent material producing reagent. Paying attention to the fact that it is in the incubation process that it must not be done, and that the luminescence reagent injection, luminescence measurement, and luminescence stop reagent injection work must be a series, the work before this incubation and the incubation The work to be performed later is separated so that each work can be performed on other containers in parallel in time.

Therefore, in the present invention, a multi-well plate having a plurality of titration vessels arranged in the XY direction,
A first driving device for driving the multi-well plate in the XY directions, and a luminescence source substance producing reagent dispensing nozzle arranged above the multi-well plate on the surface of the multi-well plate. A second drive device for driving in the X-Y directions along the same; a photometric assembly fixed above the multiwell plate for detecting the light emission of each titration container;
A photometric assembly in which a luminescence reagent injection nozzle facing the titration container and a luminescence stop reagent injection nozzle are assembled into one body, an incubation heater installed on the lower surface of the multiwell plate, and each For the titration container, injection of luminescent material producing reagent, incubation, and
A control device for separately operating the first and second drive devices at timings over all titration containers so as to achieve the five steps of injection of luminescent reagent, measurement of luminescence of titration container, and injection of luminescence stop reagent. It is characterized by

Briefly, in the conventional type, the photometric assembly and the luminescent material substance producing reagent injection nozzle are fixed on the multiwell plate, and only the multiwell plate is arranged in the XY direction. In contrast to the driving, in the present invention, the luminescence source substance generating reagent dispensing nozzle is also independent,
It is to be movable in the XY directions. That is, only the photometric assembly is fixed, and the computer is controlled so that the multiwell plate follows the movement of the reagent nozzle for producing the luminescent material and the continuous measurement is performed.

[0007]

EXAMPLE An example of the continuous titrator of the present invention will be described with reference to FIG.

A multiwell plate 5 in which a large number of titration vessels are arranged in rows and columns is placed on an XY drive table 6. A heater 11 is laid between the multi-well plate 5 and the drive table 6 over the entire surface. The multi-well plate 5 is driven in the X direction, which is the left-right direction in the figure, and in the Y direction, which is the direction perpendicular to the paper surface. Dispensing nozzle 1 in which reagent A and reagent B are mixed slightly above multiwell plate 5 from respective bottles 7 and 8 by a mixing unit 12 and injected into each of titration containers as a reagent for producing a luminescent source substance.
Which is also driven in the XY direction in a plane parallel to the driven surface of the multiwell plate 5. This drive unit is shown schematically at 16. Therefore, the dispensing nozzle 1 can select an arbitrary titration container in the multi-well plate 5 and inject the luminescent material generating reagent.

Above the multi-well plate 5, the lens unit 4 which receives the light emitted from one of the titration containers, the bottles 9 and 10 of the luminescent reagent C and the luminescence stopping reagent D, respectively, are respectively directed to the containers to be measured. There is a luminescence reagent injection nozzle 2 and a luminescence stop reagent injection nozzle 3, and these lenses and nozzles are assembled into one body to form a photometric assembly, which is fixed in a space slightly above the multiwell plate. Has been done.

The luminescence measurement by titration in one titration container will now be described in the order of its steps. One of the titration containers in which the multi-well plate is preheated by the heater 11 and the sample is put therein. First, the mixed luminescence source substance producing reagent is injected from the dispensing nozzle 1. In general, two kinds of bottles 7 and 8 are shown in the drawing because this kind of reagent dispenses an exact mixture of two reagents each time it is injected. Since the reagent B is a relatively large amount, it is introduced into the AB mixing unit 12 after being heated by the auxiliary heating unit 13. The pumping of the reagents A and B from the bottle is performed by the peristaltic pump or the tube pumps 14 and 15.

Incubation is the next step for the titration container containing the luminescent material producing reagents A and B as described above. This incubation means that the preheating is taken over and the titration container is kept at a constant temperature, but it takes a relatively long time among all the steps. As the third step, the luminescent reagent C is injected by the luminescent reagent injection nozzle 2 into the titration container after the fixed incubation time. In this case, it is preferable to jet and stir, so that the injection is performed via the syringe unit 16. As a result, the sample in the titration container starts to emit light.

The fourth step is to measure the number of photons generated by this light emission for a certain period of time. The light is taken out by a lens unit 4 arranged in the vicinity of the nozzle 2 and guided to a light receiving element 4 through an optical fiber 17, and this signal output is converted into a numerical electric signal by an electronic circuit 18, and a computer is used. Processing is performed in 19 and the result is displayed on the display 20 to be useful for analysis and research.

The light-emitting sample is stopped immediately after the photometry of the titration container is finished so that light does not leak to the adjacent titration container and does not adversely affect the next photometric operation. This step is called injection of the luminescence stopping reagent D, and the peristaltic pump 21 is supplied from the bottle 10 for the luminescence stopping reagent D.
Luminescence stop reagent nozzle 3 disposed in the photometric assembly via
More by injecting into the titration container.

FIG. 2 is a top view of the multiwell plate 5 shown in FIG. In this multi-well plate, 4 × 6 = 24 titration vessels are arranged in the XY direction. For the sake of explanation, the numbers 1 to 6 are shown in the row, and A, B, C in the column.
The symbol D is shown. This multi-well plate 5
Can be driven in the XY directions by an XY driving device as indicated by arrows XX 'and YY'. The state where the lens unit 4 is fixed above the multi-well plate 5 is shown.

The A and B reagent dispensing nozzles 1 are driven above the multiwell plate 5 in parallel with this plane by XY drive.
This is shown by the arrows of xx 'and yy', and shows the state right above the titration container A5. The above operation has been described for a series of 5 steps for one particular titration vessel, for example A1, but in the present invention it relates to how quickly a series of 5 steps is achieved for each titration vessel.

In FIG. 3, among a number of titration vessels according to the present invention,
Timechart of the steps to be taken for A1, B1, and C1
Indicates the First, the luminescence source substance-producing reagent dispensing nozzle 1 is driven above the titration container A1 to inject the reagent, and the incubation is started. Next, the dispensing nozzle 1 starts the incubation of the titration container B1 and then proceeds to the titration container C1 to start the incubation of the titration container C1. In the same manner, the operation sequence of the titration container by the driving device, that is, the stroke, is shown by an arrow line connecting the titration containers in FIG.

During this time, the incubation of the titration container A1 is completed, and the luminescent reagent C is injected from the nozzle 2 thereof, and at the same time, the measurement of the luminescence thereof is advanced by the lens unit 4. This measurement continues for a fixed time after the start of injection from the nozzle 2, and the number of photons within this measurement time is counted.
After this fixed time, the luminescence stopping reagent D is injected from the nozzle 3, and immediately after that, the multiwell plate driving device operates and the titration container B1 comes directly under the photometric assembly to perform the same work steps as described above. The titration container C1 moves with the completion of.

Looking at this as a whole, the luminescence source substance producing reagent dispensing nozzle moves to other titration containers one after another without waiting for the end of the incubation time, and the photometric assembly is set in the order of the end of the incubation time. The three-dimensional body operates so as to follow the luminescence source substance producing reagent dispensing nozzle.

This operation involves driving the two X-Y driving devices with timing, and further, the operation of each pump, the operation of opening and closing the shutter of the light receiving element, and the like, which is a computer operation. It is easily performed by the electronic control circuit 21 mainly composed of

[0020]

According to the present invention, as compared with the conventional apparatus, the incubation waiting time can be substantially effectively utilized only by adding the apparatus for driving the dispensing nozzle 1 in the XY direction. Such seed bioluminescence using a multi-well plate,
It greatly contributes to basic research of chemiluminescence, research of activity of living cells, clinical test (immunoassay), and measurement of bacterial count in food.

[Brief description of drawings]

FIG. 1 is a schematic view of a multi-well plate compatible bioluminescence measuring apparatus according to an embodiment of the present invention.

FIG. 2 is a top view of the multi-well plate shown in FIG.

FIG. 3 is a time chart showing a step work of luminescence measurement according to the present invention.

[Explanation of symbols]

 1: Luminescence source substance producing reagent dispensing nozzle 2: Luminescence reagent injection nozzle 3: Luminescence stop reagent injection nozzle 4: Lens unit 5: Multiwell plate 6: XY drive table 11: Heater 16: XY drive unit of dispensing nozzle 1 A1, B1, C1: Titration container

 ─────────────────────────────────────────────────── ─── Continued Front Page (72) Inventor Ryuji Hori 7-2-3 Hongo, Bunkyo-ku, Tokyo Ato Corporation

Claims (1)

[Claims] 1. A multiwell plate in which a plurality of titration vessels are arranged in the XY direction, and the multiwell plate is an X-plate.
A first driving device for driving in the Y direction, and a first driving device for driving the luminescent source substance producing reagent dispensing nozzle arranged above the multiwell plate in the XY direction along the surface of the multiwell plate. 2. A driving device of 2, a photometric assembly fixed above the multi-well plate for detecting the light emission of each titration container, and a luminescent reagent injection nozzle and a luminescence stop reagent injection directed to the titration container. A photometric assembly in which a nozzle and a single body are assembled, an incubation heater installed on the lower surface of the multiwell plate, and each titration container are injected with a luminescent source substance producing reagent for a predetermined time, respectively. Incubation, luminescence reagent injection, luminescence measurement of titration container,
Continuous titration luminescence measurement, characterized in that it comprises a control device for separately operating the first and second driving devices at a timing over the entire titration container so as to achieve the five steps of injection of the luminescence stopping reagent. apparatus.