JPH10108663A - Microincubator, and promotion of reaction and culture using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable agitation under the exact control of temperature that is higher than the room temperature in no need of the provision of another thermostat chamber by setting a heating chamber for the thermostat agitation equipped with the heating part surrounding the shaking bed, a fan and an openable lid on the upper face of the main body of a small-size agitator. SOLUTION: A motor 3 is installed on the center of the base plate 2 of the main body of the small-size agitator 1, and a vibration plate 6 is inserted so that it may rotate with only eccentric rotation. Between four corners of individual supporters 8 and the vibration plate 6, bars of urethane resin 9b, 9d having both flexibility and damping properties are extensively set so that the vibration plate 6 may be horizontally rotatable. A compartment 12 is set to the rear part of the upper face 1a of the main body 1 of the small size agitator and a heater 13 is attached thereto. At the face- contacting part between the front end of the machine main body 1 and the openable lid 18, approach sensors 20a and 20b for detecting the opening of the lid and stopping the actuation of' the vibration bed 11 are set to make the vibration bed 11 tentatively stop vibration, in the case that the lid is opened, even when the power source or a timer switch are not made off.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel microincubator and a method for promoting a reaction and culture using the microincubator.

[0002]

2. Description of the Related Art Conventional small stirrer (micro mixer)
Is provided with a shaking table so that it appears on the upper surface of the main body,
A microtube or microplate containing a sample solution is placed on the shaking table and shaken with stirring. Reaction tests, other reaction tests, and other culture tests in a so-called ELISA method for detecting and quantifying a target antigen or antibody using an enzyme-labeled antibody have been conventionally performed in a stationary state at room temperature.

[0003]

However, a small stirrer (micromixer) having a shaking table exposed on the upper surface of the main body cannot shake a sample solution at room temperature or at a temperature higher than the ambient temperature. There are issues. Also at room temperature,
There is a problem that the reaction or culture in a stationary state requires a long time and is still inefficient.

[0004]

SUMMARY OF THE INVENTION The present invention provides a micro-incubator provided with a heating unit, a fan, and a heating chamber for isothermal shaking, which has a heating unit, a fan, and an opening / closing lid, surrounding a shaking table on the upper surface of a small stirrer main body. Using this micro incubator, control the reaction and cultivation time of various reactions such as antibody / antigen detection, gene amplification, restriction enzymes and various culture tests of plant tissues, microorganisms, yeasts, etc., at a temperature of around 37 degrees. These problems were solved by stirring while stirring.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION

(1) Microincubator The invention of a microincubator effective for implementing the reaction / culture promotion method will be described with reference to the drawings.

A motor 3 is mounted on the center of the bottom plate 2 of the main body 1 of the small stirrer, and an eccentric rotating shaft 4 is fitted upwardly on a rotating shaft 3a projecting from the upper surface of the motor 3, and the eccentric rotating shaft 4 is mounted.
A diaphragm 6 that rotates only by eccentric rotation with the bearing 5 interposed therebetween is inserted into the side periphery of the vibration plate 6. The midsole plate 7 is laid and fixed on the bottom plate 2 of the main body 1, and the columns 8 having substantially the same height as the motor 3 are erected on the four corners of the midsole plate 7. Between 3mm and 7mm in diameter with hardness between
Urethane resin rods 9a, 9b, 9c, 9d having a hardness of about 80 to 100 and having both flexibility and vibration damping properties are stretched to support the diaphragm 6 so as to be horizontally turned. It is desirable that the diameter and hardness of the rod be determined based on a balance between required supporting force and flexibility.

Further, a balance weight 10 is attached to the eccentric rotary shaft 4 below the fitting portion of the diaphragm 6 so as to rotate synchronously.
The balance weight 10 is formed to have a weight portion 10b which spreads out in a fan shape from the fitting hole 10a in a plan view, and is fitted so that the weight portion 10b is positioned on the eccentric side. In other words, the sector 10b is positioned on the side that becomes lighter due to the eccentric rotation of the diaphragm 6, and is rotated so as to balance the weight balance, so that vibration and rattling accompanying the rotation of the diaphragm 6 are eliminated. In the drawing, a shaking table 11 that is exposed on the upper surface 1a of the main body 1 is fixedly mounted on the vibration plate 6, but the shaking table is configured such that the vibration plate 6 is exposed on the main body 1 as it is. Can also.

[0008] The compartment 12
A heater 13 is installed in the compartment 12 and the compartment
The fan 15 is attached to the perforated portion 14 provided at the upper part of the front wall 12a of the fan 12. The heater 13 is formed by laminating a heating heater 13a that is turned on / off by temperature detection and an aluminum block 13b that heats the heat generated by the heating heater 13a. Reference numeral 16 denotes an air flow hole provided in a lower portion of the front wall 12a.

A transparent opening / closing lid 18 of a vertically rotating opening / closing type which covers the front half of the upper surface 1a of the main body 1, that is, the periphery of the shaking table 11, is attached based on a hinge portion 17 provided on the upper front end of the picture chamber 12. Thus, the heating chamber 19 in which the air heated by the heater circulates is formed.

[0010] Proximity sensors 20a and 20b for detecting the opening and stopping the operation of the shaking table 11 are provided on the front end portion of the main body 1 and the contact surface formed by closing the opening / closing lid 18, and a power supply and a timer switch are provided. If the lid is opened even if it is not released, the shaking operation of the shaking table 11 is temporarily stopped.

A microtube or microplate 21 into which a reaction / culture sample solution has been dispensed is placed and fixed on a shaking table 11, the opening / closing lid 18 is closed, the power is turned on, the temperature is set, and the timer time is set. Then, the shaking table 11 is rotated and rotated by the activation of the motor 3 to stir the sample liquid, and the heater
The air heated by 13 is convected in the heating chamber 19 by the fan 15 to maintain the sample liquid at a temperature of 37 degrees.
If you want to check the state of the sample liquid on the way, when the opening / closing lid 18 is opened, the turning rotation operation is automatically stopped by the operation of the proximity sensors 20a and 20b, so that the work can be performed safely without operating each switch. . When the opening / closing lid 18 is closed, the stirring is restarted again.

(2) Method for promoting reaction in ELISA method EL using the above-described microincubator of the present invention
The reaction promoting method in the ISA method will be described as a comparison with the conventional method in the measurement test of anti-TGF beta and the result thereof.

Protocol Anti-TGF beta is diluted by a factor of two, placed in each of two microplates, adsorbed overnight at 4 ° C., and washed three times with PBS-T.

[0014] TGF-beta is diluted to 1 ng / ml and added to the microplate. The present invention uses the microincubator at 37 ° C, 700 r / min. (Humidity 33% RT).
In the conventional method, the reaction is carried out at room temperature (25 ° C., humidity 35% RT) for 2 hours each, and then the plate is washed three times with PBS-T.

A secondary antibody labeled with alkaline phosphatase is diluted 1: 5000-fold and added to a microplate. The present invention uses a microincubator at 37 ° C. and 700 r / min. (33% RT humidity). In the conventional method, the reaction is carried out for 2 hours at room temperature (25 ° C., 35% humidity RT), and the plate is washed three times with PBS-T.

A substrate solution was added to each, reacted for 10 minutes, and measured with a microplate reader.

The results of the above measurements are shown in Table 1 below. As shown in Table 1, after the addition of the enzyme-labeled antibody using the microincubator of the present invention, the reaction was promoted while stirring at a temperature of about 37 ° C. In the method, the efficiency was about 1.5 times as high as that in the conventional ELISA method which was carried out at room temperature at room temperature. Table 1 shows the average value of two tests.

[0018]

[Table 1]

In other reaction tests and culture tests, the reaction / culture time, which had been in a stationary state at room temperature, was agitated using a microincubator of the present invention at a temperature of 37 ° C. / Cultivation efficiency could be significantly improved.

[0020]

As described above, according to the present invention, the heating unit, the fan, and the heating chamber for the constant temperature shaking having the opening / closing lid are provided around the shaking table on the upper surface of the main body of the small stirrer. There is an effect that stirring under accurate temperature control higher than room temperature can be easily performed without preparing a thermostatic chamber or the like.

A proximity sensor is provided between the main body and the opening / closing lid to detect the opening of the lid and stop the operation of the shaking table. Therefore, when the lid is opened without releasing the power supply or the timer switch, the shaking is performed. The shaking of the table is temporarily stopped, and the effect is obtained that the state of the sample in the microtube or microplate set on the shaking table can be checked, or the work of taking out these can be performed easily and safely. .

The four corners of the shaking table are supported by urethane resin rods having a diameter of 3 to 7 mm, a hardness of about 80 to 100 and a flexibility and vibration-proof property. The balance weight is fitted to the eccentric rotating shaft under the shaking table, so that even if the turning rotation is repeated for a long time, the rod will be broken due to fatigue or corrosion of the rod, making it impossible to support the shaking table. In addition to the above, the vibration damping of the rod and the synchronized rotation of the balance weight can eliminate the influence of vibration on the main body due to the rotating rotation of the shaking table, thereby preventing the occurrence of noise and rattling. This produces the effect.

Using the microincubator of the present invention, the reaction / culture time is controlled while controlling the reaction / culture time for various reaction tests such as ELISA and other culture tests at a temperature of about 37 ° C. By doing
The reaction / culture efficiency at the same time can be significantly improved, or the reaction / culture time can be shortened,
There is an effect that the culture test can be performed rationally and efficiently.

[Brief description of the drawings]

FIG. 1 is a longitudinal sectional side view showing an embodiment of a micro incubator of the present invention.

FIG. 2 is a plan view showing a configuration of a rod body and a balance weight on an intermediate bottom plate.

[Explanation of symbols]

 1 is a main body 1a is an upper surface 2 is a bottom plate 3 is a motor 3a is a rotating shaft 4 is an eccentric rotating shaft 5 is a bearing 6 is a diaphragm 7 is a middle bottom plate 8 is a support 9a, 9b, 9c, 9d is a urethane resin rod 10 Is a balance weight 10a is a fitting hole 10b is a fan-shaped weight portion 11 is a shaking table 12 is a compartment 12a is a front wall 13 is a heater 13a is a heater 13b is an aluminum block 14 is a perforated portion 15 is a fan 16 is a flow hole 17 Is a hinge part 18 is an opening / closing lid 19 is a heating chamber 20a, 20b is a proximity sensor 20 is a microplate

──────────────────────────────────────────────────の Continued on front page (51) Int.Cl. ⁶ Identification code FI C12Q 1/68 G01N 33/53 T G01N 33/53 C12N 15/00 A

Claims (5)

[Claims] 1. A micro-incubator comprising a heating unit, a fan, and a heating chamber for shaking at a constant temperature having an opening / closing lid surrounding a shaking table on an upper surface of a main body of the small stirrer. 2. The microincubator according to claim 1, further comprising a proximity sensor between the main body and the opening / closing lid for detecting the opening of the lid and stopping the operation of the shaking table. 3. The four corners of a shaking table which is rotated and rotated by rotation of an eccentric rotation shaft have a thickness of 3 to 7 mm and a hardness of 80 to 100.
A urethane resin rod having a degree of hardness and flexibility and vibration-proofing is supported so as to be rotated horizontally and horizontally, and is tuned and rotated under the shaking table of the eccentric rotation shaft. The microincubator according to claim 1 or 2, wherein a weight is fitted to eliminate the influence of vibration on the main body due to the swing vibration of the shaking table. 4. Antibody / antigen detection and gene amplification in a microincubator provided with a heating unit, a fan, and an open / close lid and surrounding a shaking table on the upper surface of the main body of the small stirrer. The microincubator is characterized in that the reaction and the culturing time are controlled at a temperature of about 37 degrees C. while controlling the reaction and cultivation time of various reactions such as reaction of various enzymes, restriction enzymes, etc., and various culture tests of plant tissues, microorganisms, yeasts, etc. The reaction and culture promotion method used. 5. An enzyme-labeled antibody, for example, by binding an antigen or antibody to be measured to an antibody or antigen insolubilized on a solid phase and detecting and quantifying the antigen or antibody bound on the solid phase with the enzyme-labeled antibody. The method according to claim 4, wherein the reaction is carried out while stirring at a temperature of about 37 ° C after the addition of the enzyme-labeled antibody in the ELISA method used.