JPH11230939A - Capillary cassette and its manufacture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To reduce time for forming a window for detection, and at the same time to form the window for detection that can perform accurate detection. SOLUTION: In a manufacturing method, capillary columns 102 adhere one another for forming a line on a flat plate 204 so that a region 102a that is intended to form a window for detection is positioned on the long hole of the flat plate 204, the long hole of a flat plate 206 is placed so that it is positioned in the region 102a that is intended to form the window for detection, and the capillary columns 102 are put between the flat plates 204 and 206 for fixing. The longitudinal direction of a heated Nichrome wire coil 208 is brought into contact with the end part of the region 102a that intended to form the window for detection via the long hole of the flat plate 206 while orthogonally crossing to the capillary column 102, and a covering is burned for eliminating. The Nichrome wire coil 208 is moved along the long hole of the flat plate 206 while one part of the contact region of the Nichrome wire coil 208 is overlapped so that the covering does not remain in the region 102a that intended to form the window for detection. At this time, the covering on the surface of an opposite side where the Nichrome wire coil 208 is brought into contact is also eliminated.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field 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 fixed by a holder on a sample injection side and arranged in a line in a plane on a detection side and a method of manufacturing the same. More particularly, the present invention relates to a capillary cassette having a detection window from which a coating of a capillary column is removed in a capillary column arrangement on the detection side, and a method of forming the detection window.

[0002]

2. Description of the Related Art Electrophoresis apparatuses are used for separating and analyzing nucleic acids, proteins, peptides, sugars and the like, and play an important role in analyzing the base sequence of DNA. In order to determine the nucleotide sequence of a DNA having a long nucleotide sequence such as the human genome, an electrophoresis apparatus having high sensitivity, high speed, and large throughput is required. As one of the methods, a multi-capillary electrophoresis apparatus has been proposed in which a plurality of capillary columns filled with a gel are arranged in place of the one using a flat slab gel. Capillary columns not only allow easier handling and injection of samples than slab gels, but also allow high-speed migration and high-sensitivity detection. In other words, when a high voltage is applied to a slab gel, problems such as broadening of the band and the occurrence of a temperature gradient occur due to the influence of Joule heat.However, such a problem is small in a capillary column. Even if it does, the band spread is small and high-sensitivity detection can be performed.

[0003] Capillary columns are protected with various coatings to increase their mechanical strength. As the coating, polyimide, silicone resin, polytetrafluoroethylene, acrylic resin, or the like is used. However, when used as a coating on a capillary column for electrophoresis, optical means such as absorbance measurement and fluorescence measurement are used to detect the sample migrating in the capillary column. An obstacle to detection. Therefore, the coating on the detection unit is removed by burning with a flame using a burning means such as a lighter or a gas burner. In a multi-capillary electrophoresis apparatus that uses a plurality of capillary columns to detect a plurality of samples at the same time, a detection window is formed by aligning and fixing the capillary columns from which the coating on the detection section has been removed.

[0004]

In a multi-capillary electrophoresis apparatus, a large number of capillary columns are arranged. However, if a film is removed from the detection section on each of the capillary columns, a large amount of time is required for the operation. . In addition, depending on the coating material, in the process of removing the coating material by burning with a flame, the portion located inside the flame is completely burned and removed, but at a place away from the flame, it melts but is removed. In some cases, a projection formed by solidification of the molten coating is formed on both sides of the portion from which the coating is removed. When such convex portions are formed, the capillary columns are arranged, and when the detection window is formed by aligning the positions of the portions where the coating has been removed, the adjacent capillary columns do not adhere to each other, or because of the convex portions. The detection windows may not be aligned in a plane due to a step difference. As a result, at the time of detection, the focus of the optical system may be shifted and accurate detection may not be performed.

[0005] The present invention provides a capillary cassette having a detection window of a capillary column used in a multi-capillary electrophoresis apparatus formed in a planar shape so as to be suitable for optical measurement, and a capillary cassette characterized by the step of forming the detection window. And a method for producing the same.

[0006]

In the capillary cassette of the present invention, a plurality of capillary columns are fixed by a holder on a sample injection side and are arranged in a line in a plane on a detection side. A strip-shaped detection window extending in the capillary column arrangement direction is formed by removing the coating of the capillary column, and the capillary columns are closely adhered to each other around the detection window, and the coatings are fused together to form an integrated capillary column arrangement. Is what it is.

When manufacturing the capillary cassette, the manufacturing method of the present invention is a method for forming a detection window including the following steps (A) and (B). (A) a step of fixing the sample injection side of the capillary column with a holder, and then holding the capillary columns in close contact with each other in a planar manner and holding them in a line; (B) a capillary column in a region for forming a detection window of the held capillary column array. The heating means having a plurality of lengths is brought into contact with each other to remove the coating of the plurality of capillary columns, and the coating around the area where the coating is removed is melted and solidified to form a coating between the adjacent capillary columns. Fusing together.

[0008] Capillary columns from which no coating has been removed from the detection section are arranged and fixed in a line in a plane, and the coating is simultaneously removed from a plurality of capillary columns at positions where detection windows are to be formed by the heating means. By simultaneously heating and removing the coating of multiple capillary columns, when the molten coating cools and solidifies away from the area where the coating has been removed, adjacent capillary columns are fused and integrated. You. Thereby, a plurality of capillary columns arranged in a line form a flat cable.

[0009]

DESCRIPTION OF THE PREFERRED EMBODIMENTS As a heating means, a nichrome wire heater or a ceramic heater can be used. When a heating means having a length shorter than the width of the capillary column array on the detection side is used as a heating means for removing the coating, the step (B) is repeated plural times in the width direction of the capillary column array. A detection window is formed.

[0010]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a schematic perspective view of a capillary cassette according to one embodiment in which a plurality of capillary columns are arranged and a multi-capillary electrophoresis apparatus to which the cassette is applied. The capillary cassette 2 has a plurality of capillary columns 102 arranged therein and is fixed by holders 4 and 6 to form a capillary array. One end 2a of the capillary array is a sample injection side and is two-dimensionally formed by the holder 4. It is arranged and fixed, and comes into contact with the buffer solution of the reservoir 62 for electrophoresis after sample injection.
At the end 2b of the capillary array, the capillary columns 102 are arranged in a line, and a reservoir 56
Contact with buffer solution. At the end side of the capillary array, a detection section 2c in which the capillary columns 102 are arranged in a line and supported by the holder 6 is provided. Capillary column 102 is coated with a coating to protect it from breakage. Many capillary columns, for example, 384 capillary columns, are arranged in the capillary cassette 2. Different samples are respectively injected into each of the capillary columns 102, and electrophoresis is performed at the same time.

In the detection portion 2c of the capillary cassette 2, the coating on the capillary column 102 is removed to form a band-like detection window extending in the capillary column arrangement direction. In the vicinity of the detection window, the capillary columns 102 are in close contact with each other, and the coatings are fused together, so that the capillary column arrangement is integrated.

For example, an argon gas laser device 8 is provided as an excitation light source for exciting the sample itself or a fluorescent substance labeled on the sample. Reference numeral 10 denotes an excitation / reception optical system which irradiates the capillary column 102 of the detection section 2c with an argon laser beam of excitation light to detect fluorescence from the sample, and which is scanned by a scanning mechanism (not shown). 2c, the scanning direction (capillary column 102) parallel to the array surface of the capillary cassette and orthogonal to the electrophoresis direction.
Scanning direction). In order to prevent the excitation light beam from the laser device 8 from being shifted by the scanning of the excitation / light receiving optical system 10, the laser device 8 is used as an example here.
Is guided to an excitation / light receiving optical system 10 through an optical fiber 9 coupled by a coupler.

The lower reservoir 62 is filled with a buffer solution for electrophoresis, the lower end 2a of the capillary array is immersed in the buffer solution, and a voltage for electrophoresis is applied to the capillary end of the lower end 2a of the capillary cassette 2 through the buffer solution. Is done. The upper electrode 54 is immersed in and contacts the buffer solution of the upper reservoir 56, and the lower electrode 58 is immersed and contacts the buffer solution of the lower reservoir 62.
An electrophoresis voltage is applied from the high-voltage power supply 60 to 4,4,58.
The power supply voltage is, for example, 30 kV, and the current capacity is 10 to 3
0 mA. The sample to be electrophoresed is a protein sample, a DNA fragment sample labeled with a fluorescent substance, or the like.

One embodiment of the manufacturing method of the present invention for forming a detection window at the position of the detection target 2c in the capillary cassette 2 will be described with reference to FIG. (A) After fixing the sample injection side of the 384 capillary columns 102 from which no coating has been removed by a holder, the other end side, that is, the detection side is brought into close contact with each other, and the flat plate 2
04 in a row. In order to prevent the capillary column 102 from adhering to the flat plate 204 due to the melting of the coating by the coating removing operation, a long hole longer than the array width of the capillary column 102 is formed in the flat plate 204. Window formation scheduled area 10
2a is positioned.

(B) Another flat plate 206 is overlaid on the capillary column 102. A long hole longer than the array width of the capillary columns 102 is also formed in the flat plate 206, and the long hole is formed in the detection window formation scheduled area 102a.
Position so that it comes to. Thus, the capillary column 102 is fixed between the two flat plates 204 and 206.

(C) An electric current is applied to the nichrome wire coil 208 for burning the coating to make it glow red. The nichrome wire coil 208 is a nichrome wire 40 cm having a diameter of 0.5 mm.
Is wound into a coil having a diameter of about 3 mm and a length of about 2 cm. The amount of electricity is, for example, 5A. The longitudinal direction of the nichrome wire coil 208 is aligned with the arrangement direction of the capillary columns 102 (capillary column 102) at the end of the detection window formation scheduled region 102a through the long hole of the flat plate 206.
In a direction perpendicular to the longitudinal direction of the capillary column 10).
Touch 2

(D) After the nichrome wire coil 208 is brought into contact with the capillary column 102 for about 10 seconds to burn and remove the film at that position, the nichrome wire coil 208 is removed so that the film does not remain in the detection window forming scheduled area 102a. While partially overlapping the area where
The nichrome wire coil 208 is moved along the long hole of the flat plate 206 toward the other end of the detection window formation scheduled area 102a. At this time, the nichrome wire coil 2
The coating on the side opposite to the side where 08 contacts is also burned off.

Each of the capillary columns 102 melted at a position away from the nichrome wire coil 208 during combustion.
When the coating is cooled and solidified, the adjacent capillary columns 102 are fused together by the coating once melted.
As a result, the entire plurality of capillary columns 102 arranged in a line form a flat cable.

In the embodiment, a coiled nichrome wire is used as the heating means, but an elongated ceramic heater or the like may be used as the heating means. When the number of capillary columns fixed to the cassette holder is large, the width when the capillary columns are arranged in a line in a plane becomes large. Therefore, if the length of the nichrome wire coil is increased in order to shorten the time required for removing the coating, the strength is weakened, and the nichrome wire coil bends and deforms while the detection window is being created by pressing against the capillary column. In some cases, it is not possible to make good contact with the capillary columns arranged in a plane, and it may be necessary to repeat the combustion operation many times. In such a case, if a ceramic heater longer than the width when the capillary columns are arranged in a line in a plane is used, it is possible to form the detection window at one time, thereby shortening the detection window formation time. be able to.

[0020]

According to the capillary cassette of the present invention, since the capillary columns are closely attached to each other around the detection window and the coatings are fused together to form a capillary column array, the detection windows are aligned in a plane. The focus of the optical system at the time of detection does not deviate, and accurate detection can be performed. In the manufacturing method of the present invention, after arranging a plurality of capillary columns in a plane, the coating of the detection unit of the plurality of capillary columns is simultaneously removed, so that the detection is performed in comparison with the case where the coating is removed for each capillary column. Window creation time can be greatly reduced. In addition, at the periphery of the detection window formed by the manufacturing method of the present invention, when the coating that has been melted but not removed is cooled and solidified, the adjacent capillary columns are fused together. Since the plurality of capillary columns to be fused are arranged in a plane beforehand, the detection window is kept flat after the fusion, so that the capillary cassette of the present invention can be easily manufactured. Furthermore, when fixing the detection side of a plurality of capillary columns, the time and effort for fixing one by one can be saved, and since the fixing is not performed using an adhesive, the time for the adhesive to dry can be saved. As described above, according to the present invention, it is possible to reduce the time required for forming a detection window of a capillary column used in a multi-capillary electrophoresis apparatus and to form a detection window capable of performing accurate detection.

[Brief description of the drawings]

FIG. 1 is a schematic perspective view showing a capillary cassette of one embodiment and a multi-capillary electrophoresis apparatus to which the capillary cassette is applied.

FIG. 2 is a schematic perspective view showing a final step of an embodiment of the manufacturing method.

[Explanation of symbols]

 2 Capillary cassette 102 Capillary column 102a Expected detection window formation area 204, 206 Flat plate 208 Nichrome wire coil

Claims (3)

[Claims] 1. A plurality of capillary columns used in a multi-capillary electrophoresis apparatus are fixed by a holder on a sample injection side and are arranged in a line in a plane on a detection side. A strip-shaped detection window extending in the capillary column array direction is formed by removing the coating, and the capillary columns are in close contact with each other around the detection window, and the coatings are fused together to integrate the capillary column array. Capillary cassette characterized by the following. 2. A plurality of capillary columns used in a multi-capillary electrophoresis apparatus are fixed by a holder on a sample injection side, are arranged in a line in a plane on a detection side, and the coating of the capillary column is removed on the detection side. A method for manufacturing a capillary cassette having a detection window formed therein, comprising: forming a detection window including the following steps (A) and (B). (A) a step of fixing the sample injection side of the capillary column with a holder, and then holding the capillary columns in close contact with each other in a planar manner and holding them in a line; (B) a capillary column in a region for forming a detection window of the held capillary column array. The heating means having a plurality of lengths is brought into contact with each other to remove the coating of the plurality of capillary columns, and the coating around the area where the coating is removed is melted and solidified to form a coating between the adjacent capillary columns. Fusing together. 3. The capillary cassette according to claim 2, wherein the length of the heating means is shorter than the width of the capillary column array on the detection side, and the step (B) is repeated a plurality of times in the width direction of the capillary column array. Production method.