JP3417348B2 - Capillary column alignment device - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a plurality of capillary columns used in a multi-capillary electrophoresis apparatus, etc., which are two-dimensionally arranged on one end side and arranged in a plane in one row on the other end side. The present invention relates to a capillary column alignment device that aligns the other end side of a capillary column having one end side fixed in a planar shape when manufacturing a capillary cassette. The capillary column alignment device is applied to a capillary cassette production device that automatically produces a capillary cassette.

[0002]

2. Description of the Related Art Electrophoresis devices are used for separating and analyzing nucleic acids, proteins, peptides, sugars, etc., and particularly play an important role in analyzing the base sequence of DNA. DNA with a long nucleotide sequence like the human genome
In order to determine the nucleotide sequence of (1), an electrophoretic device having high sensitivity, high speed, and large processing capacity is required. As one of the methods, a multi-capillary electrophoresis apparatus in which a plurality of gel-filled capillary columns are arranged instead of using a flat slab gel has been proposed. Compared with slab gel, the capillary column is not only easier to handle and inject a sample, but also allows high-speed electrophoresis and high-sensitivity detection. In other words, if a high voltage is applied with a slab gel, there are problems such as band broadening and temperature gradient due to the effect of Joule heat, but with a capillary column, there are few such problems, and high voltage is applied for high-speed electrophoresis. Even if it is turned on, the band spread is small and high-sensitivity detection is possible.

In a multi-capillary electrophoresis apparatus for simultaneously detecting a plurality of samples, a plurality of capillary columns are fixed to a holder on the sample introduction side and the detection side and mounted in the apparatus in the state of a capillary cassette. A plurality of capillary columns are arranged in the capillary cassette, and the capillary array is configured by being fixed by the sample introduction side holder and the detection side holder, and the plurality of capillary columns are constituted by the sample introduction side holder and the detection side holder. For example, on the sample introduction side, they are arranged two-dimensionally in a matrix, and on the detection side, they are arranged in a plane in one row.

The capillary cassette is automatically manufactured by the capillary cassette manufacturing apparatus. FIG. 1 is a schematic perspective view showing a conventional method of manufacturing a capillary cassette. In FIG. 1, the number of capillary columns is reduced and shown. A sample introduction side holder 6 for arranging the sample introduction side ends of the capillary column 4 is provided. The holder 6 has, for example, 16 × 24 holes with a pitch of 4.5 mm, and the sample introduction side end of the capillary column 4 is inserted into each of these holes to form a two-dimensional array. Holder 6
Are moved in the XY plane by the holder moving mechanism of the capillary cassette manufacturing apparatus. Capillary column 4
Alignment groove 5 having a width dimension slightly larger than the outer diameter of the capillary column 4 in order to align the detection side ends of the two in a row in a plane.
Is provided with a slit holder.

When the capillary cassette is manufactured, the capillary column 4 is pulled out from the drum (not shown) around which the capillary column 4 is wound along the X-axis in the drawing, and the end portion which is the sample introduction side end is held by the holder 6. After leading to the hole, the capillary column 4 is cut into a predetermined length. The hole position (address) of the holder 6 into which the sample introduction side end is inserted is selected by moving the holder 6 in the YZ plane. The end (detection side end) on the cut side of the capillary column 4 is dropped into the alignment groove 5. Since the detection-side end is aligned in a plane in the order of being dropped into the alignment groove 5, the detection-side end can be aligned corresponding to the address of the holder 6.

[0006]

However, as in the conventional example shown in FIG. 1, if one capillary column is inserted into each of the many holes formed in the holder 6, it will take a long time to manufacture the capillary cassette. It takes time. Therefore, in order to reduce the manufacturing time of the capillary cassette, as shown in FIG. 2, there is provided a capillary cassette manufacturing apparatus capable of simultaneously inserting one row or one column of capillary columns into the holes of the holder 6 arranged in the Y-axis direction. Conceivable. In this case, in order to align the detection side of the capillary column 4 with the alignment groove 5 corresponding to the address of the holder 6, after inserting a plurality of capillary columns into the holes of the holder 6 aligned in the Y-axis direction at the same time, 6 is moved in the Y-axis direction and dropped into the alignment groove 5, the capillary column 4 is moved onto the alignment groove 5, cut into a predetermined length, and dropped into the alignment groove 5 for one line. Alternatively, it is possible to repeat for one column of the number of capillary columns, but this results in a large time loss, and the benefit of simultaneous insertion is not much received. Up to this point, the capillary cassette used in electrophoresis has been described, but the present invention is not limited to this, and flexible capillaries are arranged two-dimensionally on one end side and flat on the other end side. This applies to all capillary cassettes.

According to the present invention, a plurality of capillary columns are two-dimensionally arranged on one end side, and a capillary column array capable of shortening the production time of a capillary cassette in which one end side is arranged in a line in a plane is arranged. The purpose is to provide a device.

[0008]

According to the present invention, when a plurality of capillary columns are two-dimensionally arranged on one end side and one end is planarly arranged in a single row, a capillary cassette is manufactured. A capillary column alignment device for aligning the other end side of a capillary column whose one end side is fixed in a plane, wherein the capillary column is sequentially dropped, and an alignment groove for aligning the other end side of the capillary column in a plane shape, A belt on the surface of which a plurality of recesses for accommodating one row or one column of the other end of a capillary column arranged in a matrix on one end side is formed, and the belt is rotated. A belt rotating mechanism for sequentially dropping the capillary columns housed in the recesses into the alignment grooves in an order according to the position of the arrangement on one end side thereof.

After one end side of one row or one column of capillary columns is simultaneously fixed to a predetermined array position by the capillary cassette manufacturing apparatus, the other end side of the capillary columns is adjusted according to the position of the array on the one end side. And store them one by one in the recess of the belt. Then, the belt is rotated by the belt rotating mechanism to convey the other end side of the capillary column to the alignment groove side, and the capillary columns housed in the recesses are sequentially dropped into the alignment groove in the order according to the arrangement position. Thereby, even if one end side of one row or one column of the column is arranged at the same time, the other end side of the capillary column can be aligned in a plane according to the arrangement position.

It is preferable that the recesses of the belt are formed at an arrangement interval on one end side of the capillary column or at an interval which is an integer fraction of the arrangement interval. As a result, one end side of a plurality of capillary columns that are arranged in the same row or column of the arrangement on one end side of the capillary column is arranged at the same time, and when the other end side is cut off, the other capillary columns are The end side can be accurately accommodated in the recess. here,
When the recesses of the belt are formed at intervals of an integer fraction of the arrangement interval on one end side of the capillary column, the capillary column is not housed between the capillary columns on the other end side housed in the recesses and between the capillary columns. There is an empty recess. For example, the arrangement interval on one end side is 9 mm, and the interval between the concave portions of the belt is ½ of the arrangement interval on the one end side.
When formed with 5 mm, the other end of one row or one column of capillary columns is accommodated in every other recess.
Also, using the belt, the arrangement interval on one end side is 4.5 m.
The other end side of the capillary cassette of m can also be aligned, so that versatility can be provided.

[0011]

EXAMPLE FIG. 3 is a schematic perspective view of a multi-capillary electrophoresis apparatus as an example of application of a capillary cassette manufactured by using the apparatus of the present invention. The capillary cassette 2 has a plurality of capillary columns, for example, 384 capillary columns 4 arranged therein, and is fixed by a sample introduction side holder 6 and a detection side holder 8 to form a capillary array. The outer diameter of the capillary column 4 is 0.3 m
m.

One end 2a of the capillary array serves as the sample introduction side, and a holder 6 is used to measure 16 × 2 of 4.5 mm pitch.
4 are arranged and fixed in two dimensions, and come into contact with the buffer solution for migration contained in the reservoir 10 for migration after the sample injection. At the end (detection side end) 2b of the capillary array, the capillary columns 4 are aligned in one row and come into contact with the buffer solution contained in the reservoir 12.
One capillary column 4 is provided on the side of the capillary array end 2b.
A detection window 2c arranged in a row and supported by the holder 8 is provided. The capillary column 4 is coated with acrylic or polyimide to protect it from damage, but the coating is removed from the detection window 2c. Different samples are injected into the respective capillary columns 4 and simultaneously electrophoresed.

As the excitation light source 14 for exciting the sample or the fluorescent substance labeling the sample, for example, an argon gas laser device is provided. Reference numeral 16 denotes an excitation / reception optical system, which irradiates the detection window 2c with a beam of excitation light,
The fluorescence from the sample is detected, and the fluorescence is scanned by a scanning mechanism (not shown) in the scanning direction parallel to the array surface of the capillary array in the detection window 2c and orthogonal to the migration direction. In order to prevent the pumping light beam from the pumping light source 14 from being displaced by the scanning of the pumping / receiving optical system 16, here, as an example, the laser beam from the pumping light source 14 is pumped through an optical fiber 18 coupled by a coupler. It is guided to the light receiving optical system 16.

A buffer solution is contained in the reservoir 10,
The capillary array end 2a on the sample introduction side is immersed in the buffer solution, and a migration voltage is applied to the capillary column end of the capillary array end 2a through the buffer solution.
A buffer solution is contained in the reservoir 12, the detection-side capillary array end 2b is immersed in the buffer solution, and a migration voltage is applied to the capillary column end of the capillary array end 2b through the buffer solution. The upper electrode 22 is immersed in and in contact with the buffer solution in the reservoir 12, and the lower electrode 20 is immersed in and in contact with the buffer solution in the reservoir 10,
A migration voltage is applied from a high voltage power supply 24 to both electrodes 20, 22. The power supply voltage is, for example, 30 kV, and the current capacity is 10 to 30 mA. The sample to be electrophoresed is a protein sample or a DNA fragment sample labeled with a fluorescent substance.

Next, an embodiment of the present invention will be described. This embodiment comprises an alignment groove unit and a support unit. 4 to 7 are schematic configuration diagrams showing the alignment groove unit, FIG. 4 is a front view, FIG. 5 is a top view, and FIG.
Is a left side view, and FIG. 7 is a right side view showing the vicinity of the fixed air source nozzle. A pair of arm members 26, 26 forming a frame
Are provided facing each other. Rollers 28 and 30 are arranged at both ends of the upper side surfaces of the arm members 26 and 26, respectively, and belts 32 are provided on the rollers 28 and 30, respectively. One capillary column is installed on the outer peripheral surface of the belt 32.
Recesses 34 for accommodating books one by one are formed (see the detailed view of the broken line circle portion in FIG. 6). The distance between the centers of the adjacent recesses 34 is 4.5 mm.

Rollers 36, 36, whose fixing positions can be changed, are provided to tension the belts 32, 32. Sprockets 38, 38 having grooves formed to match the intervals of the recesses 34 for rotating the belts 32, 32
And a motor 40 for simultaneously rotating the sprockets 38, 38. The belt rotating mechanism of the present invention includes rollers 28, 30, 36, a sprocket 38, and a motor 40.

Alignment groove forming members 42, 42 are detachably attached to the front side surfaces of the arm members 26, 26 with screws 44, 44. Arm member 26 and groove forming member 42
Alignment grooves 46, 46 having a width of 0.38 mm are formed between and. In order to surely drop the capillary column 4 carried to the groove 46 side by the belt 32 into the groove 46, one of the aligning groove forming members 42 is supplied with air supplied from a fixed air source (not shown). A fixed air source nozzle 48 that jets downward is provided near the roller 28 (see FIG. 7). The arm members 26, 26 are mounted on the cylinder 50 fixed to the capillary cassette manufacturing apparatus, on the shaft 49.
Has been installed through. The alignment groove unit is moved up and down by the cylinder 50. The alignment groove unit is
In the capillary cassette manufacturing apparatus, a plurality of capillary columns are arranged below the capillary column position when the sample introduction side is fixed by being pulled out from the drum.

8 to 10 are schematic configuration diagrams showing the support unit. FIG. 8 is a front view, FIG. 9 is a top view, and FIG.
Is a left side view. The support unit has a width dimension between the arm members 26, 26 of the alignment groove unit,
Swap prevention plate 52 having a flat surface on the surface facing 26
Is provided. The prevention plate 52 is installed on a cylinder 56 fixed to the capillary cassette manufacturing apparatus via a frame 54 and a shaft 55. The support unit is moved up and down by the cylinder 56. The support unit is a position facing the capillary cassette alignment unit with the capillary column sandwiched above the capillary column position when a plurality of capillary columns are pulled out from the drum and the sample introduction side is fixed in the capillary cassette manufacturing apparatus. It is located in.

The frame 54 has three movable air source nozzles 58 for ejecting downwardly the air supplied from a movable air source (not shown) to the plane including the alignment grooves 46, 46 of the alignment groove unit. Are arranged. The nozzle 58 is fixed to the frame 54 so as to be arranged at a position corresponding to the bottom of the groove 46 when the capillary cassette alignment unit is raised and the support unit is lowered when aligning the capillary columns.

FIG. 11 is a front view showing the alignment groove unit and the support unit when aligning the capillary columns. FIG. 12 is a side view showing the positional relationship between the belt and the swap prevention plate when aligning the capillary columns. The operation at the time of aligning the capillary columns will be described with reference to FIGS. 4 to 12. After the capillary column 4 is pulled out from the plurality of capillary column drums by the capillary cassette manufacturing apparatus and the sample introduction side capillary column ends arranged in the same row of the sample introduction side end are fixed, the capillary column 4 is aligned by the cylinder 50. Raise the groove unit to move the cylinder 56
To lower the support unit. Swap prevention plate 52
Are arranged between the belts 32, 32 on the arm members 26, 26 when aligning the capillary columns. At this time, the swap prevention plate 52, as shown in FIG.
The flat surface of the prevention plate 52 on the side of the arm member 26 is lower than the outer peripheral surface of the belt 32 and the groove 46 is formed so that the capillary column 4 housed in 4 does not jump and move to another recess 34 when the belt 32 rotates. Is arranged between the belts 32, 32 so as to be located above the bottom of the belt.

The detection side of the plurality of capillary columns 4 is
As shown in FIG. 12, the belts 32 are accommodated one by one in the concave portion 34. At this time, the interval between the adjacent recesses 34, 34 is 4.5 mm in accordance with the arrangement interval on the sample introduction side, so that one capillary column 4 can be accurately accommodated in each recess 34. After that, the capillary column 4 is cut into a predetermined length by the capillary cassette manufacturing apparatus, and the detection side end is manufactured.

The belt 40 is rotated by rotating the motor 40, and the detection side of the capillary column 4 is conveyed to the groove 46 side. At this time, the prevention plate 52 is arranged so that the plane of the prevention plate 52 on the arm member 26 side is located below the outer peripheral surface of the belt 32 and above the bottom of the groove 46.
Since the capillaries 4 are arranged between them, it is possible to prevent the capillary column 4 housed in the recess 34 from jumping and moving to another recess 34 when the belt 32 rotates.

The capillary column 4 conveyed to the groove 46 side
Are dropped into the groove 46 away from the belt 32. At this time, by ejecting air downward from the fixed air source nozzle 48, the capillary column 4 separated from the belt 32 can be reliably dropped below the groove 46,
It is possible to prevent the capillary column 4 from being clogged in the upper portion of the groove 46. The capillary columns 4 housed in the recesses 34 are sequentially dropped into the grooves 46 in the order according to the arrangement on the sample introduction side. After aligning the detection sides of all the capillary columns 4 housed in the recesses 34 in the grooves 46, the motor 40 is stopped and the rotation of the belt 32 is stopped.

Next, the cylinder 50 lowers the alignment groove unit and the cylinder 56 raises the support unit. By ejecting air from the movable air source nozzle 58, the capillary column 4 dropped into the groove 46 is moved in order from the capillary column 4 housed in the lower portion of the groove 46. Since it is pushed downward by the air ejected from 58, the capillary columns 4 can be aligned in the groove 46 without any gap. After that, the capillary column manufacturing apparatus fixes the end of the capillary column in the next row of the sample introduction side array, and then aligns the detection sides of the capillary columns 4 in the same manner as the above operation.

[0025]

In the capillary column alignment device of the present invention, the capillary columns are sequentially dropped, and the alignment grooves for aligning the other end side of the capillary column in a plane shape and the capillary columns arranged in a matrix on one end side. A belt having a plurality of concave portions for accommodating the other ends of the capillary columns for one row or one column, and a belt rotating mechanism for rotating the belt. Since the capillary columns housed in the recesses are sequentially dropped into the alignment groove in the order according to the arrangement position, the other end side of the plurality of capillary columns whose one end side is fixed is belted according to the position of the arrangement. After accommodating them one by one in the concave part, the belt is rotated by the belt rotating mechanism, and the capillary columns accommodated in the concave part are sequentially arranged in the alignment groove in the order according to the position of the arrangement on the one end side. By inserting, even if one end side of multiple capillary columns is arranged at the same time, the other end side of the capillary columns can be aligned in a plane according to the arrangement position, and the time required for manufacturing the capillary cassette is reduced. It can be shortened. Further, by forming the concave portions of the belt at the arrangement intervals on the one end side of the capillary column or at intervals of an integer fraction of the arrangement interval, a plurality of columns arranged in the same row or column on the one end side arrangement of the capillary column are formed. The other end of the capillary column can be accurately accommodated in the recess one by one.

[Brief description of drawings]

FIG. 1 is a schematic perspective view showing a conventional method of manufacturing a capillary cassette.

FIG. 2 is a schematic perspective view showing a method of manufacturing a capillary cassette in which a plurality of capillary columns are simultaneously inserted.

FIG. 3 is a schematic perspective view showing a multi-capillary electrophoresis apparatus as an example of application of a capillary cassette manufactured by using the apparatus of the present invention.

FIG. 4 is a schematic configuration front view showing an alignment groove unit that constitutes an embodiment.

FIG. 5 is a schematic configuration top view showing the unit.

FIG. 6 is a left side view of a schematic configuration showing the unit.

FIG. 7 is a right side view of a schematic configuration showing the vicinity of a fixed air source nozzle of the unit.

FIG. 8 is a front view showing a support unit that constitutes an embodiment.

FIG. 9 is a schematic configuration top view showing the unit.

FIG. 10 is a schematic configuration left side view showing the unit.

FIG. 11 is a front view showing an alignment groove unit and a support unit when aligning a capillary column.

FIG. 12 is a side view showing a positional relationship between a belt and a swap prevention plate when aligning a capillary column.

[Explanation of symbols]

4 capillary columns 26 Arm member 28, 30, 36 rollers 32 belt 34 recess 38 sprockets 40 motor 42 Alignment groove forming member 44 screws 46 Alignment groove 48 Fixed air source nozzle 49,55 shaft 50,56 cylinders 52 Swap prevention plate 54 frames 58 Movable air source nozzle

─────────────────────────────────────────────────── ─── Continuation of the front page (58) Fields surveyed (Int.Cl. ⁷ , DB name) G01N 27/447

Claims (3)

(57) [Claims] 1. A plurality of capillary columns are connected to each other at one end side.
Capillary column alignment device for aligning the other end side of the capillary column having the fixed one end side in a plane shape when manufacturing the capillary cassettes which are arranged dimensionally and arranged in a line in a planar manner at the other end side In the above, the capillary columns are sequentially dropped, an alignment groove for aligning the other end side of the capillary column in a plane shape, and one of the capillary columns arranged in a matrix on the one end side.
A belt having a plurality of recesses formed on the surface for accommodating the other ends of the column or row columns of capillary columns one by one, and rotating the belt to accommodate the capillary columns accommodated in the recesses at the one end side. And a belt rotating mechanism that is sequentially dropped into the alignment groove in an order corresponding to the position of the arrangement. 2. The capillary column alignment device according to claim 1, wherein the recesses of the belt are formed at an array interval on the one end side of the capillary column or at an interval of an integer fraction of the array interval. 3. The capillary column alignment device according to claim 1, wherein the capillary cassette is used in a multi-capillary electrophoresis device, and the one end side is a sample introduction side and the other end side is a detection side.