JP3850153B2 - Capillary cassette for multi-capillary electrophoresis device - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a capillary cassette in which a plurality of capillary columns used in a multi-capillary electrophoresis apparatus are bundled.
Such a capillary cassette is mounted on a multi-capillary electrophoresis apparatus and is used in clinical fields, analysis fields, inspection fields, etc., and particularly used for DNA sequence analysis.
[0002]
[Prior art]
The electrophoresis apparatus is used for separating and analyzing nucleic acids, proteins, peptides, sugars, and the like, and particularly plays an important role in analysis of DNA base sequences. In order to determine the base sequence of a DNA having a long base sequence such as the human genome, an electrophoresis apparatus with high sensitivity, high speed and high throughput is required. As one of the methods, a multi-capillary electrophoresis apparatus in which a plurality of capillary columns filled with a gel (which will be referred to as a polymer because it is composed of a polymer) will be proposed instead of using a flat slab gel. ing. Capillary columns are not only easier to handle and inject samples than slab gels, but also can be migrated at high speed and detected with high sensitivity. In other words, if a high voltage is applied with a slab gel, problems such as band expansion or temperature gradients occur due to the effect of Joule heat, but such problems are few with capillary columns. Even if it is made, it is possible to perform highly sensitive detection with little band spread.
[0003]
In a multi-capillary electrophoresis apparatus that uses a plurality of capillary columns and simultaneously detects a plurality of samples, the capillary columns are fixed to a cassette holder on the sample introduction side and the detection side and installed in the apparatus. With the cassette holder, the capillary columns are two-dimensionally arranged on the sample introduction side, for example, and arranged in a line on the detection side.
[0004]
When a capillary column is filled with a polymer, one end of the capillary column is fixed by piercing the capillary column with an elastic material such as a rubber stopper, or fixed to a dedicated holder for filling the polymer and filled with a gap with an adhesive. Fix the elastic material or dedicated holder to the container containing the polymer so that the end of the capillary column is immersed in the polymer, seal the inside of the container, pressurize the inside of the container with a pump, and press the polymer into the capillary column, or An elastic material or a dedicated holder is connected to the pump, the other end of the capillary column is immersed in the polymer, the inside of the capillary column is depressurized by the pump, the polymer is sucked into the capillary column, and the polymer is filled in the capillary column.
[0005]
[Problems to be solved by the invention]
When filling the capillary column by press-fitting or sucking the polymer, the method in which the capillary column is pierced and fixed to the elastic material is insufficient in pressure resistance, and the capillary column cannot be filled well.
Also, in the method of fixing the capillary column to the dedicated holder for filling the polymer with an adhesive, the polymer cannot be filled well into the capillary column due to insufficient filling of the adhesive, and the production yield is poor.
[0006]
Accordingly, an object of the present invention is to provide a capillary cassette capable of reliably filling a polymer in all capillary columns with a high yield.
[0007]
[Means for Solving the Problems]
The present invention relates to a capillary cassette in which a plurality of capillary columns used in a multi-capillary electrophoresis apparatus are bundled, one end of which is bundled in a cylindrical shape by a sleeve, and a gap between the sleeve and the capillary and between the capillaries Is sealed with a filler.
[0008]
One end of a capillary column bundled in a cylindrical shape by a sleeve has a cylindrical outer shape, so that it can be easily installed in a polymer packing device while maintaining airtightness, and it is easy to pack a polymer using high pressure into the capillary column. become.
DETAILED DESCRIPTION OF THE INVENTION
[0009]
The sleeve is preferably made by shrinking a shrinkable member, and more preferably a heat shrinkable tube.
By passing one end of a plurality of capillary columns through a sleeve made of a shrinkable member and then shrinking the sleeve, the capillary column ends can be bundled in the most dense and cylindrical shape. If a filler is applied in advance to the capillary column surface at a position corresponding to the shrinkable member, the filler can be filled in the gap between the capillary columns without failure in the bundling process. When a heat shrinkable tube is used as the sleeve, the capillary column ends can be bundled only by heating using a dryer or the like.
[0010]
It is preferable that a mounting member for mounting on the polymer filling apparatus is attached on the sleeve while maintaining airtightness with the sleeve.
Capillary column ends bundled in a cylindrical shape by a sleeve can be handled in the same way as, for example, liquid chromatograph piping, so if a mounting member such as a ferrule is attached on the sleeve when the polymer is packed into the capillary column, the capillary column end is airtight. It can be fixed to the polymer filling device with good performance.
[0011]
It is preferable that a detection target portion in which capillary columns are arranged in a row is provided on the end portion side bundled in a cylindrical shape, and that the capillary column is two-dimensionally arranged on the other end side to serve as a sample injection portion.
By aligning the capillary column end on the other side opposite to the end side bundled in a cylindrical shape two-dimensionally, the capillary column end is immersed in various sample solutions at the time of sample injection, and the sample is simultaneously loaded into each capillary column. Can be injected. In addition, after sample injection, electrophoresis voltage can be applied to both ends of all capillary columns to simultaneously separate and detect samples in each capillary column.
[0012]
【Example】
FIG. 1 is a perspective view showing a capillary cassette of one embodiment in which a plurality of capillary columns are arranged.
A plurality of capillary columns 2 are arranged, a sample injection side is fixed by a cassette holder 4, and a detection side is fixed by a cassette holder 6 and a heat shrinkable tube 10 at an end portion to constitute a capillary cassette 8. One end 2a of the capillary cassette 8 serves as a sample injection portion and is two-dimensionally arranged and fixed by the cassette holder 4. On the other end side of the capillary cassette 8, the capillary column 2 is arranged and fixed in a line in a plane by a cassette holder 6, and the capillary column end 2 b is bundled in a cylindrical shape by a contracted heat shrink tube 10. The cassette holder 6 is formed with a detection window, and the capillary column in the detection window serves as the detected portion 2c.
The outer diameter of the capillary column 2 is, for example, 300 to 400 μm.
[0013]
FIG. 2 is a schematic diagram showing a procedure for bundling capillary column ends with a heat-shrinkable tube. 3A and 3B are cross-sectional views taken along line AA ′, and FIG. 3B is a cross-sectional view taken along line BB ′. However, the number of capillary columns is shown reduced.
(A) After aligning the plurality of capillary columns 2, a filler 12 such as an epoxy resin adhesive or silicon compound is applied to the surface of the capillary column 2 at a predetermined position from one end 2b.
[0014]
(B) The capillary column end 2b is inserted into the heat shrinkable tube 10 while being bundled, and the heat shrinkable tube 10 is put on the position where the filler 12 is applied.
(C) The heat shrinkable tube 10 is heated to shrink. The inner diameter of the heat-shrinkable tube 10 is reduced, the capillary columns 2 are in close contact with each other, and the gap is sealed with the filler 12 so as to be airtightly bundled into a cylindrical shape. The gap between the heat shrinkable tube 10 and the capillary column 2 and between the capillary columns 2 and 2 in the heat shrinkable tube 10 is sealed with a filler 12. The end side does not penetrate.
[0015]
FIG. 4 is a schematic diagram showing a method for fixing a mounting member to a polymer filling device to the end of the capillary column.
The capillary column 2 bundled by the heat shrinkable tube 10 is inserted into the ferrule 16 as a mounting member to the polymer filling device together with the heat shrinkable tube 10 and fixed by the screw 18 so that the pressure resistance at the time of injecting the polymer into each capillary column 2 is sufficient. It is a thing.
[0016]
By adjusting the outer diameter of the capillary column, the number of capillary columns, and the thickness of the heat-shrinkable tube after shrinkage, the outer size of the heat-shrinkable tube 10 after shrinkage in the state where the capillary column is inserted is changed to the outer dimension of the piping of the liquid chromatograph, for example It is preferable to set it to φ1.6 (outer diameter 1.6 mm), φ2, φ3, and the like. As a result, an existing ferrule used for a liquid chromatograph or the like can be used. In addition, when preparing a dedicated ferrule, a capillary column may be bundled according to the inner diameter of the dedicated ferrule.
In addition, since the unevenness of the bundled capillary columns is eliminated depending on the thickness of the heat shrinkable tube, it is more preferable that the heat shrinkable tube used has a larger thickness after shrinkage.
At the time of polymer filling, this ferrule 16 is connected to and attached to the connecting portion of the polymer filling device, and the polymer is filled into the capillary column by press-fitting or suction.
[0017]
FIG. 5 is a schematic perspective view showing an embodiment of a multi-capillary electrophoresis apparatus to which the capillary cassette of FIG. 1 is applied. In FIG. 5, the cassette holder is omitted.
A pair of reservoirs 110 and 120 accommodates electrophoresis buffer solutions 112 and 122, respectively, and electrodes 130 and 132 are provided in both buffer solutions, respectively.
[0018]
A sample is accommodated in each well 102 of the sample plate 100. A wiring pattern is formed on the sample plate 100, and electrodes are inserted into the wells 102. The electrodes are connected to the high-voltage wiring cable via the connector portion 106.
The reservoir 110 and the sample plate 100 are connected so as to be switched by a high-voltage switching unit 136 so that the wiring is switched, and an electrophoresis high-voltage power source 134 is connected between the high-voltage switching unit 136 and the electrode 132 provided in the other reservoir 120. The voltage for sample injection and the voltage for electrophoresis are switched and applied between them.
[0019]
At the time of sample injection, one end 2a of the capillary column constituting the capillary cassette 8 is inserted into each well 102 of the sample plate 100 one by one, and after sample injection is switched to the reservoir 110 and the one end 2a is immersed in the buffer solution 112. The The other end 2 b of the capillary cassette is immersed in the buffer solution 122 of the other reservoir 120 together with the heat shrinkable tube 10.
An optical measurement unit 14 that detects a sample by absorbance or fluorescence is provided in the vicinity of the detected portion 2c.
One of the sample plate 100 and the reservoir 110 is selectively switched by a moving mechanism (not shown in FIG. 1) so as to contact the capillary column end 2a.
[0020]
Next, the operation of this embodiment will be described.
At the time of sample injection, the capillary column end 2 a is immersed one by one in the well 102, and the capillary column end 2 b is immersed together with the heat shrinkable tube 10 in the buffer liquid 122 of the reservoir 120. Then, the high voltage switching unit 136 is connected to the sample plate 100 side, and a high voltage is applied between the well 102 and the reservoir 120 by the high voltage power supply 134 for electrophoresis. The sample in the well 102 is injected into the capillary column 2.
[0021]
After the sample injection, the high voltage application is temporarily stopped, and the sample plate 100 and the reservoir 110 are moved by the moving mechanism, so that the capillary end 2a on the sample side is immersed in the buffer solution 112 of the reservoir 110. Thereafter, electrophoretic separation is performed by applying a high voltage between the reservoirs 110 and 120. The voltage for injecting the sample into the capillary column and the power supply voltage for electrophoresis are, for example, 30 kV, and the current capacity is 10 to 30 mA.
The separated sample components sequentially pass through the detected portion 2c, and are detected by the optical measuring portion 14 at that time.
In such a multi-capillary electrophoresis apparatus, an automatic polymer filling mechanism is provided, the used polymer in the capillary column is discharged by the automatic polymer filling mechanism, and the capillary column is reused so as to be filled with a new polymer. It is preferable.
[0022]
【The invention's effect】
In the capillary cassette of the present invention, one end thereof is bundled in a cylindrical shape by a sleeve, and the gap between the sleeve and the capillary and between the capillaries is sealed with a filler. For example, it can be handled in the same manner as the piping of a liquid chromatograph, and when the polymer is packed into the capillary column, it can be hermetically fixed using a connecting member such as a ferrule. The polymer can be filled.
[Brief description of the drawings]
FIG. 1 is a perspective view showing a capillary cassette of one embodiment in which a plurality of capillary columns are arranged.
FIG. 2 is a schematic diagram showing a procedure for bundling capillary column ends with a heat-shrinkable tube.
3A and 3B are views showing a cross section of the end of the detection-side capillary column of the embodiment, where FIG. 3A is a cross-sectional view along the line AA ′, and FIG. 3B is a cross-sectional view along the line BB ′ is there.
FIG. 4 is a schematic cross-sectional view showing a state where a mounting member to a polymer filling device is fixed to the end of the capillary column.
FIG. 5 is a schematic perspective view showing an embodiment of a multi-capillary electrophoresis apparatus to which the embodiment is applied.
[Explanation of symbols]
2 Capillary column 2b Capillary column end 10 Heat shrinkable tube 12 Packing agent

Claims (4)

In a capillary cassette in which a plurality of capillary columns used in a multi-capillary electrophoresis apparatus are bundled,
One end thereof is bundled in a cylindrical shape by a sleeve, and a gap between the sleeve and the capillary and between the capillaries is sealed with a filler,
A capillary cassette, wherein a mounting member for mounting on a gel filling device is mounted on the sleeve while maintaining airtightness between the sleeve and the sleeve.   The capillary cassette according to claim 1, wherein the sleeve is formed by shrinking a shrinkable member.   The capillary cassette according to claim 2, wherein the shrinkable member is a heat shrinkable tube.   4. A detection part in which capillary columns are arranged in a row is provided on the end side bundled in a cylindrical shape, and the capillary column is two-dimensionally arranged on the other end side to form a sample injection part. The capillary cassette according to any one of the above.

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