JPH02143145A - Electrophoresis device - 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 an electrophoresis device. More specifically, the present invention relates to an electrophoresis device that uses light and has a dark room that can maintain a constant temperature of an electrophoresis plate.

[Prior Art] Gel electrophoresis is widely used as a method for determining the base sequence of DNA and the like.

Conventionally, samples were labeled with radioisotopes and analyzed during electrophoresis, but this method had the disadvantage of being laborious and time-consuming. For this reason, recently, a method of labeling the sample with a fluorescent material is being considered.

In the method using light, fluorescently labeled DNA fragments are electrophoresed in a gel, and from the start of the electrophoresis, 15 to 20 CIB
A photoexcitation unit and a photodetector are provided for each migration path at the bottom.
The DNA fragments that pass through this point are measured in order. The detection time determines the electrophoresis speed, ie, 1) the size of the NA fragment.

JP-A No. 63-21556 discloses a structure in which the line of gel 1- of an electrophoresis device irradiated with a laser and the array direction of the photodiode array are perpendicular to the direction of migration of DNA fragments in the electrophoresis device. A DNA jtf base sequencing device is disclosed.

FIG. 3 is a schematic diagram illustrating the configuration of the device.

As shown in the figure, the laser light emitted from the light source 40 is bent by a mirror 42 and irradiates the migration plate 44 from the lateral direction. When the fluorescent label DNA 46 reaches the irradiation area, it emits fluorescence. Fluorescent light from each migration path is imaged by the lens 48 on the light receiving section 52 of the image intensifier 50. This signal is amplified and converted into an electrical signal by a photodiode array 54, which is then used as an electrical signal.

[Problems to be Solved by the Invention] A migration plate used for gel electrophoresis has a gel-like electrolyte layer interposed between two quartz or glass plates.

When vertical gel electrophoresis is actually carried out using an apparatus such as that disclosed in Japanese Patent Application Laid-Open No. 63-21556, the electrophoresis plate is housed in a dark room that can be sealed, and a laser beam is irradiated from the side. As measurements are continued in this manner, the temperature in the dark room rises by -1- due to the heat of the laser beam, and the temperature of the migration plate also rises at the same time.

Electrophoresis must be performed while keeping the temperature of the electrophoresis plate constant. If the temperature of the electrophoresis plate is unstable, not only will the measurement results be adversely affected, but the reliability of the δ111 constant will also be impaired.

In order to keep the temperature of the electrophoresis plate constant, conventional methods used water cooling of the electrophoresis plate. However, this method had many problems in terms of performance and maintenance, and there was a strong desire to develop another constant temperature method.

Therefore, an object of the present invention is to provide an electrophoresis device that uses laser light and does not use water as a means for keeping the temperature of the migration plate constant.

[Means for Solving the Problems] As a means for achieving the above-mentioned object, the present invention provides an airtight housing in which a migration plate is accommodated and a stage for irradiating the migration plate with laser light. In an electrophoresis measurement apparatus which has a darkroom with a function of 1000 kHz, a Ntl'l memorizing darkroom symbol chamber Kyrene-type means is provided to keep the temperature of the electrophoresis plate and the darkroom constant.

Circulation r stage has air outlet and υj′ air outlet
A fan is attached to at least one of the air outlet, air outlet, and exhaust outlet.

Air blower 1-1 is placed directly above the laser beam irradiation stage, and JJ
It is preferable that the I air outlet is disposed facing the air outlet.

It is preferable that an intake duct bent in a crank shape is connected to the feeder JJI [1, and an air duct bent in a crank shape or r air duct is connected to the υ1-wind [1].

[Function] As described above, in the electrophoresis apparatus of the present invention, the temperature of the electrophoresis plate and the temperature inside the dark room can be maintained constant by providing a circulation means in the dark room and ventilating the air in the dark room. can.

Unlike conventional water-cooling systems, the circulator system is extremely easy to maintain because it only requires air to flow. In addition, in the case of the circulation method, the /&J degree of the electrophoresis plate and the dark room can be easily controlled by simply changing the amount of air electricity being sent.

In the case of a water cooling system, this can be handled by changing the sprayed water n1, but there is a limit to the increase in water π. Also, san/li
If the water meter is increased, the 4 degrees on the electrophoresis plate can be lowered within a short time, but a relatively long response time is required to lower the temperature in the darkroom. This makes temperature control complicated.

In the present invention, since the temperature of the electrophoresis plate and the temperature inside the dark room are approximately the same, temperature control is relatively easy. For example, in a water-cooled system, the temperature of the electrophoresis plate and the temperature inside the darkroom may differ significantly, so temperature sensors must be installed in both the electrophoresis plate and the darkroom, but in the present invention, the ambient temperature is determined by the temperature of the electrophoresis plate. Therefore, the temperature sensor only needs to be installed in the dark room, and there is little need to attach the temperature sensor to the electrophoresis plate.

In the case of a water cooling system, there are problems with corrosion and rust of the device, but according to the circulation system of the present invention, such problems do not occur at all. For this reason, the installation j27. maintenance becomes extremely easy.

[Example] Hereinafter, the electrophoresis apparatus of the present invention will be described in more detail with reference to the drawings.

FIG. 1 is a conceptual diagram of a darkroom portion of the electrophoresis apparatus of the present invention, and FIG. 2 is a perspective view of the darkroom with the door open.

As shown in FIG. 1, a migration plate 3 is placed within the chamber. A laser beam irradiation end 5 is provided on a side wall at a position perpendicular to the electrophoresis plate 3, and a laser beam 7 is irradiated from this to the electrophoresis plate 3 in the lateral force direction. The temperature near the laser beam irradiation end 5, especially directly above it, tends to rise by 42 degrees due to the heat emitted from the laser beam. For this reason.

Laser light! ! (It is preferable to provide an air outlet 9 at the direct line 1- of the radiation end 5. A side wall facing the side wall on which the air outlet is arranged, on the same axis as the air outlet, that is, facing this air outlet. It is preferable to provide an air outlet 11 at the side of the electrophoresis plate 3. With such a configuration, the air introduced into the dark room 1 from the air outlet 9 flows while touching the side surface of the migration plate 3, so that the air does not draw a straight line of flow. υ1 wind] flows into 111.

If outside light enters the dark room from ventilation LI and ventilation [1], there is a risk of measurement errors, so in order to prevent outside light from entering, , an intake duct 13 and an exhaust duct 15 bent into a crank shape are attached.

In order to circulate air in the darkroom, it is necessary to connect an air blower or a uF air fan to at least one of the intake duct and the jJF air duct. In a preferred embodiment, the blower 119 includes a blower fan 17.
Directly attached the haifu fan 19 to the exhaust II 11.
Attach directly.

As shown in FIG. 2, a door 17i'+21 which can be opened and closed is disposed in front of the darkroom 1, so that the darkroom can be hermetically sealed during measurements.

The electrophoresis plate 3 is held directly by a electrophoresis plate holder 23 in a dark room. Buffer tanks 25 and 27 for storing J7 solution, which is necessary for gel '1u aerophoresis, are provided in one part and the lower part of the holder 23. The first end and the lower end of the electrophoresis plate are immersed 711t in the buffer solution in each buffer 7/tank. 1. In order to conduct electricity from the L buffer solution to the lower buffer solution through the gel electrolyte layer between the electrophoresis plates.

Electrode 3 is installed in the lower buff γ tank and the lower 11 Cubanofa box, respectively.
0 and 32 are provided. The electrophoresis plate 3 can be attached to and detached from the holder 23 by the clamp mechanism 34 .

As is clear from FIG. 2, a laser beam irradiation end 5 is provided on the side wall at a position perpendicular to the migration plate 3. Apply the laser beam so that the optical axis of the laser beam irradiation end matches the gel electrolyte layer of the electrophoresis plate! ! (A radiation end 5 is provided. As shown in the figure, an air outlet or a ventilation fan is provided directly to the laser beam radiation end 5. Opposite this air outlet, the side wall on the opposite side is provided with an outlet 34 for the exhaust duct.

The type of opening/closing door 21 is not limited to the sliding door type shown. You can use any type that can seal the darkroom, such as the double door type and the flip-up type.

[Effects of the Invention Part 1] As explained above, in the electrophoresis apparatus of the present invention, a circulation L stage is provided in the darkroom, and the temperature of the electrophoresis plate and the inside of the darkroom are controlled by ventilation of the air in the darkroom. The temperature
can be maintained at a constant level.

Unlike conventional water cooling systems, the Circulane and In systems are extremely easy to maintain because they only allow air to flow. Further, in the case of the circulation method, the temperature of the electrophoresis plate and the dark room can be easily controlled by simply changing the supplied air [11].

In the case of a water cooling system, this can be handled by changing the sprayed water jl, but there is a limit to the increase in water M. In addition, spray water 1
4, the temperature of the electrophoresis plate can be lowered within a short time, but it takes a relatively long response time to lower the temperature inside the dark room. This makes l&1 degree control complicated.

In the present invention, the temperature of the electrophoresis plate and n7? Since the indoor temperature is similar to that of the human body, temperature control is relatively easy.

For example, in a water-cooled system, the temperature of the electrophoresis plate and the temperature inside the dark room may differ significantly, so it is necessary to immerse a temperature sensor in both the electrophoresis plate and the dark room, but in the present invention, the ambient temperature is Since it represents temperature, the 714 degree sensor only needs to be installed in a dark room, and there is little need to attach a temperature sensor to the electrophoresis plate.

In the case of a water cooling system, there are problems with corrosion and rust of the device, but according to the circulation system of the present invention, such problems do not occur at all. Therefore, maintenance of the device becomes extremely easy.

[Brief explanation of the drawing]

FIG. 1 is a conceptual diagram of the darkroom section of the electrophoresis apparatus of the present invention, FIG. 2 is a perspective view of the darkroom with the door open, and FIG. 1 is a schematic configuration diagram of a DNA base sequencing device disclosed in . 1... Dark room, 3... Electrophoresis plate, 5... Laser light irradiation end. 9...Air blowing 0.11...Air exhaust [7] 13...Intake duct. 15...υ1 air duct.

Claims (4)

[Claims] (1) An electrophoresis measuring device having a hermetically sealed dark room in which a migration plate is housed and a means for irradiating the migration plate with laser light, characterized in that a circulation means is provided in the dark room. electrophoresis device. (2) The electrophoresis device according to claim 1, wherein the circulation means has an air outlet and an air outlet, and a fan is attached to at least one of the air outlet and the air outlet. (3) The electrophoresis device according to claim 1 or 2, wherein the air outlet is disposed directly above the laser beam irradiation means, and the air outlet is disposed facing the air outlet. (4) An intake duct bent in a crank shape is connected to the air outlet, and an exhaust duct bent in a crank shape is connected to the air outlet. Electrophoresis apparatus described.