JPH0624768Y2 - Improved electrophoresis device - Google Patents

Description

[Detailed description of the device] (a) Field of industrial application Recently, in the field of experimental research in the fields of medicine, pharmacy, agriculture and genetic engineering, analysis of protein or nucleic acid molecules has become popular. As a method of analysis, electrophoresis tends to be particularly often used because of its advantage that electrophoresis is relatively safe and accurate.

In view of this, the present invention adds effective technical means in order to eliminate the drawbacks of the conventional method such as the leakage of the chemical solution in the gel container or the smile phenomenon in the migration result of the apparatus used for electrophoresis, and its advantages The present invention relates to an improved electrophoretic device capable of exhibiting properly, and an object thereof is to obtain an electrophoretic device suitable for industrialization.

(B) Conventional technology Conventionally, a vertical slab electrophoresis method has been used in a method for analyzing a molecule of protein or nucleic acid. If an example of an instrument used for this method is shown in FIGS. The vertical slab electrophoresis apparatus as shown in FIG. 5 is described below.

First, agarose or polyacrylamide gel of a polysaccharide is mainly used as a medium for analyzing a protein or nucleic acid molecule in this type of device. Further, as shown in FIG. 3, the gel 7 is provided with spacers 19 at the left and right ends of the inner glass plate 16 and the outer glass plate 15 so that a thin space is formed therein so that the gel 7 can be accommodated therein. As shown in FIGS. 4 and 5, 28 is closely attached by the clip 20 and is still fixed to the upper buffer tank 25 by the clamp 17.

Then, a buffer solution containing an electrolyte such as Tris is used for guiding the current to the gel 7, but from the anode terminal 14, the current is applied to the gel 7 through the upper buffer solution 6 filled in the upper buffer tank 28. Further, a current is led from the cathode terminal 18 to the gel 7 through the lower buffer solution 13 in the migration tank 8. For example, when an analytical sample containing a protein is placed on the uppermost part of the gel 7 and direct current is applied from the anode to the cathode, the charged protein starts to migrate toward the lower cathode.

In this case, since the migration speed varies depending on the property of the protein, the protein naturally causes a phenomenon of being separated according to the type.

Therefore, if you look at the condition, turn off the current, take out gel 7, and stain only the protein, it will be separated into stripes by type, so you can confirm that they have been separated, and run them in parallel with the standard sample. For example, it is said that protein molecules can be identified, and quantification can also be performed depending on the staining concentration.

[References]

Published by Ito Dental Publishing Co., Ltd. Hideo Asakawa Kiyoko Sano Co-authored Clinical Laboratory Lecture P69-78 (c) Problems to be solved by this device (a) Into the glass sandwich shown in Fig. 3, for example, When molding a polyacrylamide gel, the acrylamide monomer may leak, but even if some of the monomer leaks in this way, the gel shape will change and it can be used for electrophoresis. It disappears, but I try to solve this point. As seen in the glass sandwich of FIG. 2, since the gel is filled up to the lower end of the bottom, it deforms due to the elasticity of the packing rubber and becomes unusable, but this point will also be addressed.

(b) In the above-mentioned glass sandwich, the gel and the upper buffer solution are in contact with each other with the glass plate in between, so there is a dislike that the temperature distribution of the gel during electrophoresis cannot be kept uniform. An attempt is made to solve the problem that a phenomenon occurs and an appropriate migration result cannot be obtained.

(c) In the electrophoretic tank shown in FIG. 2, during the electrophoresis, the buffer solution leaked from the upper buffer solution tank at a time, and the amount of the buffer solution was reduced, or the buffer solution was completely lost and the electrophoresis was interrupted. There is a risk that the migration tank will burn out, but we will try to solve this point.

(D) Means for solving the problem (a) U-shaped cassette plate 4 having a U-shaped spacer 30 on the front surface and a connection port 9 on the lower rear surface as shown in FIGS. 12 and 14. A gel cassette 27 is used in which a plastic film 5 is horizontally ultrasonically welded to the surface of a spacer 30 to form a thin space for the gel 7 in which the central portion of the plate is completely sealed.

(b) As shown in FIG. 13, the surface of the U-shaped spacer 30 of the gel cassette 27 is inward on the vertical surface of the packing rubber 11 embedded in the same shape on the front and rear sides of the U-shaped cassette holder 12. A method is adopted in which the inside of the hollow portion thus formed is closely contacted with the upper buffer solution tank 25 filled with the upper buffer solution 6.

(c) As shown in FIG. 7, insert the cassette holder 12 in which the gel cassettes 27 are closely attached to the front and rear sides of the electrophoresis tank 8 and continue backward to strongly press the gel cassettes 27. The pressing device 2 and the crimping plate 3 as shown in FIG. 15 are installed, and the vertical pressure generated by pressing the upper plate of the pressing device 2 is set so that the slopes of both sides compete with each other. Adopts a crimping tool that converts into lateral pressure and acts as lateral pressure.

(E) Action (a) As shown in FIG. 10, the gel cassette 27 is a thin type formed by forming the front surface of the cassette plate 4 into a U-shaped spacer 30 and flatly welding the plastic film 5 on the surface. Since the space is made into a gel 7 container, it serves to completely prevent liquid leakage, and since it is protected by the U-shaped spacer 30 of the gel cassette 27, it serves to prevent deformation of the gel 7. .

(b) As can be seen from FIGS. 12 to 14,
The gel cassette 27 has a function of adhering to both sides of the cassette holder 12 to form an upper buffer solution tank 25, and the one-sided gel cassette 27 allows the upper buffer solution 6 and the gel 7 to pass through the plastic film 5. To make contact with each other sufficiently.

(c) As seen in FIG. 15, when the pusher 2 is pushed down into the space 26, a pressure in the direction of the arrow M is generated, and the pressure plate 3 is pressed.
Receives the pressure and converts the pressure into parallel pressure in the direction of the arrow N, and this serves to press the gel holder 12 fitted in the front part of the migration tank 8 under pressure. Further, the two setscrews 21 provided on the rear upper surface of the pressing device 2 are for maintaining the above-mentioned crimping action, and the return screw 1 provided in the middle thereof adjusts the pressurization or presses the pressing device 2. The action of pulling out.

(F) Example (i) First, a gel cassette 27 as shown in FIGS. 12 and 14 was prepared.

This gel cassette 27 has a U-shaped spacer 30 portion having a 1 cm width and a 1 mm thick outer edge portion on the front surface, and a deformation preventing groove 10 of substantially the same shape as that on the back surface, which is formed in parallel with the upper portion of the bottom of the groove. 2 mm wide and 8 cm wide connection port 9 for steady current flow
Injection molding of the cassette plate 4 having a U-shaped spacer 30
A plastic film 5 having a thickness of 0.38 mm, which had been subjected to pickling on the inner surface, was ultrasonically welded horizontally to the surface so that the thin hollow part of the gel 7 container having a length and width of 7.8 cm square was contained.

(ii) Next, the cassette holder 12 was prepared as shown in FIG.

This cassette holder 12 uses a Bakelite plate with a thickness of 1 cm and a width of 1.8 cm to make a U-shaped device with a bottom of 8 cm and sides of 11.5 cm, and at the center of each of the front and back sides,
Anode wire 23 in which a rectangular packing rubber 11 for closely contacting the gel cassette 27 is embedded in a U shape and is wired in the inner space
And a single terminal 22 for connecting and energizing both of the cathode lines 24 wired to the outside of the right side is projected to the outside of the right side.

(iii) Next, as shown in FIG. 15, the pusher 2 and the crimping plate 3
One set of crimping tools was prepared.

These are rectangular rectangular box-shaped bodies of similar shape obtained by cutting a rectangular body with a length of 10 cm, a width of 5 cm, and a height of 10 cm along a diagonal line on the front surface. The left part is the pressing device 2 and the other is a crimping plate. 3, the cassette holder 12 with the gel cassette 27 of the object to be pressured on both sides is installed in the front part of the migration tank 8, and the crimping plate 3 is placed in contact therewith, and finally the slope of the pressing device 2 is stepped. When the device is joined and intervened and the device is pressed, the crimping plate 3 competes laterally and the object to be pressured comes to be crimped. It was constructed so that only three parts were in contact with the surface of the U-shaped spacer 30.

(iv) The equipment prepared as described above was loaded into the electrophoresis tank 8 to complete the improved electrophoresis apparatus.

The electrophoretic tank 8 was a solid box-shaped rectangular parallelepiped having a vertical and horizontal depth of 11 cm each by adhering a Bakelite plate having a thickness of 0.5 to 1.0 cm.

Further, as shown in FIG. 7, in the same tank, the cassette holder 12 having the upper buffer tank 25 formed by closely adhering the gel cassettes 27 on the front and the back at the front is placed, and then the crimping plate 3 is attached. Then, the pressing device 2 was press-fitted, and the cassette holder 12 was pressure-bonded to the cassette holder 12. After that, the fixing screw 21 was installed so that the cassette holder 12 could be maintained. Thus,
The lower buffer solution 13 was injected into the migration tank 8 and the upper buffer solution 6 was injected into the upper buffer solution tank 25, the sample was placed on the gel 7, and current was passed from the single terminal 22 to start migration. . As a result of examining the repeat performance of this trial, it was proved that it has a very effective function for solving the problems described above.

(G) Effect of the device Although it has been generally called electrophoresis, which has many failures, the present invention has established and utilized it as a safe and reliable analysis method, and has a high reproducibility of results and a reliable method. It was shown to be very effective.

The following points are the winning effects.

(a) Since a completely sealed gel container was provided in the center of the gel cassette and a deformation preventing groove was provided on the back surface, the concern about liquid leakage of the gel container and distortion or deformation of the gel was eliminated.

(b) The back side of the gel cassette is provided with a connection port, and a flat thin plastic film is welded on the front side, so that the current flows evenly, and the gel and the buffer solution are sufficiently contacted, The temperature distribution was kept uniform and the smile phenomenon disappeared, and the migration results showed a straight striped band.

(c) By providing a crimping tool to strongly crimp the gel cassette, there is no fear of liquid leakage from the upper buffer tank.

[Brief description of drawings]

FIG. 1 is an overall explanatory view of the present invention by the AA cross section, FIG. 2 is an explanatory view of a conventional method illustrated, FIG. 3 is an explanatory view of the gel, FIG. 4 is an explanatory view of using the clamp, and FIG. Is an explanatory view of using the clip, FIG. 6 is a front view of the present invention, FIG. 7 is a plan view thereof, FIG. 8 is a front view of the same gel cassette, FIG. 9 is a sectional view of the same EE, and FIG. 10 is the same. Plan view, FIG. 11 is a sectional view of the same FF, FIG.
The figure shows the gel cassette front perspective view, FIG. 13 shows the cassette holder rear perspective view, FIG. 14 shows the gel cassette rear perspective view, and FIG.
FIG. 16 is a structural view of the present invention taken along the line BB, FIG. 16 is a perspective view of a pressing device, and FIG. 17 is a perspective view of a crimping plate. Explanation of symbols 1 ... Return screw, 2 ... Presser, 3 ... Crimping plate, 4 ... Cassette plate, 5 ... Plastic film, 6 ... Upper buffer solution, 7 ...
Gel, 8 ... migration tank, 9 ... connection port, 10 ... deformation prevention groove, 11 ...
Packing rubber, 12 ... Cassette holder, 13 ... Lower buffer solution, 14 ... Anode terminal, 15 ... Outer glass plate, 16 ... Inner glass plate, 17 ... Clamp, 18 ... Anode terminal, 19 ... Spacer, 20
… Clip, 21… Set screw, 22… Single terminal, 23… Anode line, 24… Cathode line, 25… Upper buffer tank, 26… Void part, 27
… Gel cassette, 28… Glass sandwich, 29… Notched part, 30… U-shaped spacer

Claims (1)

[Scope of utility model registration request] 1. A gel cassette 27 comprising a U-shaped spacer 30 on the surface of a cassette plate 4 and a plastic film 5 welded thereto.
An improved electric device characterized in that the cassette holder 12 forming the upper buffer tank 25 by closely adhering to each other and the pressing device 2 and the crimping plate 3 are fitted in a rectangular electrophoretic tank 8. Electrophoretic device.