JPH0561583B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an electrophoresis device, and particularly to a large-sized electrophoresis device suitable for sample separation.

[Conventional technology]

The conventional device is as described in Japanese Patent Application Laid-Open No. 59-52743.
The structure was such that the electrophoresed sample was collected through a collection hole along with a buffer solution. However, no consideration was given to the flow velocity distribution of the bath liquid in the direction of the bath thickness and the accompanying differences in the migration distances of components of equal charge.

[Problem that the invention seeks to solve]

The above-mentioned prior art does not take into account the differences in the flow velocity distribution in the tank thickness direction and the resulting migration distance in the thickness direction, and the flow velocity near the tank wall is low and the migration time and migration distance are long. Even though the flow rate of the central part is high and the migration distance is short, the samples were collected all at once from the collection holes at the same distance, so there was a problem that samples of equal charge could not be separated well.

An object of the present invention is to provide an electrophoresis device that can separate samples of equal charge with high precision.

[Means for solving problems]

To achieve the above object, the present invention provides an electrophoresis apparatus in which a sample injection tube and a buffer introduction tube are provided at the upper part of an electrophoresis tank having a rectangular cross section, and electrodes are provided at both ends in the width direction. The electrophoresis tank is provided with a plurality of sample collection holes in the center in the thickness direction, and a tank liquid discharge hole for discharging unseparated tank liquid from these sample collection holes is located downstream of the sample collection hole. It is characterized by the fact that it is provided.

[Effect]

According to the above configuration, the flow velocity distribution in the tank thickness direction is parabolic, with zero at the tank wall and maximum at the center, with a small velocity difference at the center and a large velocity difference near the tank wall.
Therefore, if the sample collection holes are arranged in the center of the tank, liquid with a small speed difference will flow in, and the remaining liquid near the tank wall with a large speed difference will be discharged from a discharge hole provided downstream.
When the speed difference between the fractionated liquids is small, the electrophoretic distances of the samples of equal charge become equal, so that the samples of equal charge can be fractionated with high accuracy from the fractionation holes of the same distance.

〔Example〕

An embodiment of the present invention will be described below with reference to FIG. The main components are an electrophoresis tank 1, a sample injection tube 2,
Preparation section 3, buffer inlet tube 4, tank liquid discharge tube 5, - electrode 6, + electrode 7, membranes 8, 9, sample injection pump 10,
Tank liquid pump 11, preparative pump 12, preparative tube 13, preparative container 14, fixing plate 15, rectifying plate 16,
Sample container 17, buffer solution introduction container 18, tank solution discharge container 19, electrolyte solution tank 20, separation hole 21, migration tank bottom 22, tank solution discharge hole 23, tube groove 24, holding plate 25, taper 26, shikiri plate It is 27. Also,
The system is composed of a sample system 101, a buffer system 102, a bath liquid system 103, and a preparative sample system 104.

The electrophoresis tank 1 has a buffer introduction tube 4, a sample injection tube 2 at the top, a - electrode 6, a + electrode 7 at both ends, and the electrophoresis tank 1.
This member includes an electrolytic solution tank 20 that is in contact with the electrolytic solution tank 20 through membranes 8 and 9, a separation section 3 at the bottom, and a tank liquid discharge pipe 5 at the bottom. The buffer introduction tube 4 is a member for introducing the buffer solution 102 into the migration tank 1, the sample injection tube 2 is a member for injecting the sample 101 into the migration tank 1, and the fractionation section 3 has a collection hole 21 at the top and a collection hole at the bottom. A member including a tube 13, fixing plates 15 at both ends, a rectifying plate 16 on the side wall, and a buffer solution discharge pipe 5. The tank liquid discharge pipe 5 is a member for discharging the tank liquid 103 that is not fractionated from the sorting hole 21. , tank liquid pump 11
is for introducing the buffer solution 102 from the buffer solution introduction container 18 into the migration tank 1 through the buffer solution introduction pipe 4, and introducing the tank liquid 103 in the liquid movement tank into the tank liquid discharge container 19 through the tank liquid discharge pipe 5. The sample injection pump 10 is a member that injects the sample 101 from the sample container 17 into the migration tank 1 through the sample injection tube 2, and the preparative pump 12 is a member that injects the sample 104 separated by electrophoresis from the preparative hole into the preparative tube 13. This is a member for introducing the sample into the preparative container 14 through the .

The electrodes 6 and 7 are members for applying an electric field to the tank liquid in the migration tank 1, and the fixing plate 15 and the rectifying plate 16 are fixed so that the separation hole 21 is arranged at the center of the migration tank 1 in the thickness direction. It is also a member for making the flow in the migration tank 1 in the separation hole 21 parallel.

Buffer solution 102 is introduced into sample 10 from buffer solution introduction tube 4.
1 is introduced into the electrophoresis tank 1 through the sample injection tube and an electric field is applied to the tank liquid using the electrodes 7 and 8. Since the electrophoresis distance varies depending on the strength of the charge, the sample can be separated into its components. The separated preparative sample 104 is separated into the preparative container 14 through the preparative hole 21 . Further, the uncollected tank liquid 103 is introduced into the tank liquid discharge container 19 from the tank liquid discharge pipe 5 at the bottom of the migration tank 1.

2 is a side view of the embodiment of FIG. 1; FIG. When the buffer solution 102 is allowed to flow down from the top of the electrophoresis tank 1, the flow velocity distribution in the thickness direction of the electrophoresis tank 1 is a parabolic shape with the maximum at the center and 0 at the wall of the tank. The smaller the electrophoresis distance, the smaller the variation in the migration distance of components with equal electric charge. Therefore,
The sample 101 is allowed to flow down from the center in the thickness direction of the electrophoresis tank 1, subjected to electrophoresis, separated from the collection holes 21 arranged in the center of the electrophoresis tank 1, and the remaining tank liquid 103 is passed through the tank liquid discharge pipe at the bottom of the electrophoresis tank. It was discharged from 5.

According to this embodiment, the separation holes are arranged in the center in the thickness direction of the electrophoresis tank and the sample is separated, so that components having equal charges can be separated with high precision. Further, according to this embodiment, since the rectifier plate is provided downstream of the separation hole and the tank liquid discharge hole is provided downstream of the separation hole, there is an effect of obtaining a parallel flow in the separation hole.

FIG. 3 is a detailed view of the sorting section shown in FIG. 1, in which figure a is a plan view, figure b is a front view, and figure c is a side view. A preparative hole 21 is arranged at the tip of the preparative part 3, and a preparative tube 13 is arranged below it, and a migration tank bottom 22, a tank liquid discharge hole 23, and a tank liquid discharge pipe 5 are provided in the lower tank wall. The structure is such that the separation section 3 is sandwiched between the walls and the separation hole 21 is fixed at the center in the thickness direction of the electrophoresis tank.

In addition, in this embodiment, the thickness of the preparative section 3 is made thin so that the sample is collected only from the center in the thickness direction of the electrophoresis tank 1. The separation hole 21 is designed to reduce the pitch of the separation hole 21 so that as much of the electrophoretically separated sample as possible can be collected.
The shape of is a rectangle with a large cross-sectional area, and the preparative hole 21
In order not to disturb the flow in the electrophoresis tank, a taper 26 is provided at the tip, and a filter plate 2 between the separation holes 21 is provided.
7 was made thinner.

FIG. 4 is a detailed view of the sorting hole 21 shown in FIG. 3. The sorting hole 21 is constructed by providing a large number of concave grooves in two plates and gluing them together at the slit plate 27.

FIG. 5 shows how the separation tubes 13 are arranged. The preparative tube 13 is sandwiched between the tube grooves 24 of a pair of presser plates 25, which have a tube groove 24 on one side, a migration tank bottom 22, and a tank liquid discharge hole 23 on the other side, and the three are glued together for separation. The sorting section 3 is constructed by arranging the sorting tubes 13 and providing the member for the aforementioned sorting hole 21 at the tip.

According to this example, the pitch between the preparative holes is small;
Moreover, since the entrance shape of the separation hole is rectangular and a thin separation section can be constructed, it is possible to provide a separation section with a high sample separation effect without disturbing the flow in the electrophoresis tank.

[Effects of the Invention] According to the present invention, since a sample can be separated only from the center in the tank thickness direction where the difference in velocity of the tank liquid in the electrophoresis tank is small, it is possible to separate samples with equal charges with high precision. be.

[Brief explanation of drawings]

Fig. 1 is a front view of one embodiment of the present invention, Fig. 2 is a side view of Fig. 1, Figs. 3 a to c are detailed views of the sorting section of Fig. 1, and Fig. 3 a is a plan view. , FIG. 3b is a front view, FIG. 3c is a side view, FIG. 4 is a detailed view of the sorting hole, and FIG. 5 is a detailed view of the sorting tube arrangement. DESCRIPTION OF SYMBOLS 1...Migration tank, 2...Sample injection tube, 3...Separation section, 4...Buffer solution introduction tube, 5...Bath liquid discharge tube, 6
...-electrode, 7...+electrode, 8,9...membrane, 10
... Sample injection pump, 11 ... Tank liquid pump, 12
...Preparative pump, 13...Preparative tube, 14...
…Preparation container, 15…fixing plate, 16…straightening plate,
17...sample container, 18...buffer introduction container, 1
9... Tank liquid discharge container, 20... Electrolyte tank, 21...
...Preparation hole, 22... Bottom of electrophoresis tank, 23... Tank liquid discharge hole, 24... Tube groove, 25... Holding plate, 2
6... Taper, 27... Shikiri plate, 101... Sample system, 102... Buffer system, 103... Tank drainage system, 104... Preparation sample system.

Claims (1)

[Claims] 1. In an electrophoresis device in which a sample injection tube and a buffer introduction tube are provided at the upper part of an electrophoresis tank having a rectangular cross section, and electrodes are provided at both ends in the width direction, the electrophoresis tank has a rectangular cross section in the center in the thickness direction. A plurality of sample sampling holes are provided, and a tank liquid discharge hole for discharging uncollected liquid from these sample sampling holes is provided on the downstream side of the sample sampling hole. Electrophoresis device.