JP2001174467A - Sample container for biochemical analysis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sample container for biochemical analysis which is capable of efficiently treating a large number of samples and can be treated for sterilization, etc., at a high temperature and repeatedly used. SOLUTION: Through the use of a sample container 1 for biochemical analysis in which a plurality of recessed parts 2 are formed at pitch intervals W of 500 μm or less on the surface of a plate-shaped body made of glass and/or ceramic, biochemical analysis such as DNA analysis is performed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sample container for biochemical analysis which is suitably used for DNA analysis such as genetic diagnosis and gene determination for determining a nucleic acid (DNA) sequence.

[0002]

2. Description of the Related Art In recent years, various analytical techniques have been known in the field of biochemistry, such as a DNA analysis method in which a nucleic acid (DNA) sequence is analyzed to make a genetic diagnosis or a genetic determination. For example, in a hybridization method which is an example of a DNA analysis method, a DNA probe having a specific DNA sequence is immobilized on the surface of a support such as beads or a glass plate,
The support and the buffer solution are placed in a container made of a resin such as polystyrene or Teflon, and a sample solution having DNA is injected into the container to perform hybridization to determine whether complementary binding with the specific DNA occurs. After performing the operation, the solution is cultured, and the presence or absence of a specific DNA is determined by observing whether or not a predetermined DNA has proliferated.

Conventionally, in this type of analysis, Japanese Patent Application Laid-Open
As disclosed in JP-A-505729, a plurality of test tubes are arranged and each DNA probe and a sample droplet are dropped in the test tubes, or the surface of a substrate divided by a grid-like partition wall of 1 to 10 mm is used. The above-mentioned DN is provided on the inner surface of each section.
This has been done by dropping a plurality of droplets (spots) of several microns containing the A probe, dropping a sample droplet onto the DNA probe by pipetting, culturing, and observing.

[0004]

However, in the analysis of genetic diagnosis and the like, if a conventional individual container is used to handle an enormous number of samples, the operation is troublesome, or the volume of the container increases once. In addition, there is a problem that a large amount of sample cannot be processed, and when a plurality of spots are placed on a plane in one section as described above, there is a possibility that droplets may be mixed due to vibration or the like during work, and There was a problem that the interval between the samples could not be narrowed and the sample container was enlarged.

In a conventional container made of an organic resin,
Heating at high temperature and high pressure causes deterioration, so sterilization at high temperature and high pressure cannot be performed, sample containers are delicate, and cannot be reused by heating after use. there were.

[0006] The present invention has been made to solve the above problems, and can efficiently process a large amount of samples.
An object of the present invention is to provide a biochemical analysis container that can be sterilized at high temperature and high pressure and that can be used repeatedly.

[0007]

As a result of studying the above-mentioned problems, the present inventors have formed a plurality of recesses on the surface of a plate made of glass and / or ceramic as a sample container for biochemical analysis such as DNA analysis. It has been found that by using a container made of such a material, a large amount of sample can be efficiently processed, a sterilization treatment at high temperature and high pressure can be performed, and the container for DNA analysis can be used repeatedly.

That is, the container for biochemical analysis of the present invention comprises:
A plurality of recesses are formed at a pitch of 500 μm or less on the surface of a substrate made of glass and / or ceramic, and the recesses are formed in a groove shape, or partition walls between the recesses are formed in a grid shape. Is desirable.

In addition, the elution amount of metal ions after filling the concave portion with water and holding for 1 hour is 1 ppm or less.
Further, it is desirable that the content of the alkali metal in the glass and / or ceramic is 5% or less.

The container for biochemical analysis of the present invention can be suitably used especially for DNA analysis.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS An example of a sample container for biochemical analysis of the present invention will be described with reference to a schematic sectional view of FIG. FIG.
In the above, a sample container for biochemical analysis (hereinafter, simply referred to as a container) 1 has a plurality of recesses 2 formed on a surface of a substrate made of glass and / or ceramics.

The material of the container 1 is quartz glass, soda lime glass, low soda glass, lead silicate glass, borosilicate glass, lead glass, alkali silicate glass, bismuth glass, etc., alumina, zirconia. , Silica, magnesia, and other ceramics can be suitably used, and it is particularly desirable to contain glass in view of cost and ease of production. Furthermore, improvement of mechanical strength, heat resistance and alkali metals such as Na, K, Li and Mg, C
From the viewpoint of suppressing the deterioration of DNA due to elution of metal components such as a, Sr, and Ba, low-soda glass containing less sodium and lead is desirable.

When a DNA probe is used as a DNA analysis technique by a hybridization method, it is desirable that a hydroxyl group is supported on at least the inner wall surface of the container 1, and at least the inner wall surface of the container 1
It is desirable to contain the i component, particularly to contain amorphous silica glass. Thus, the hydroxyl group on the inner wall surface of the container 1 can be chemically bonded to the DNA probe, so that the DNA probe can be easily carried on the inner wall surface of the container 1, and the container 1 serves as a support for fixing and holding the DNA probe. Can be combined.

Further, in the container 1, whiteness is improved.
, Coloring, increasing the strength of the container 1, etc.
Therefore, the average particle size of SiO_Two, ZrO_Two,
Al _TwoO_Three, TiO_Two, Si_ThreeN_Four, Fe_TwoO_Three, Ni_TwoO_Three,
At least one filler such as CuO, MnO, BN, etc.
It can also be added.

Further, in order to improve the dimensional accuracy at the time of manufacturing,
The container 1 may be formed of a material different from the material of the substrate 3 forming the bottom surface of the concave portion 2 and the material of the partition wall 4 forming the side wall surface of the concave portion 2. In this case, the softening temperature of the material of the substrate 3 forming the bottom surface of the concave portion 2 is changed to the partition wall 4 forming the side wall surface of the concave portion 2.
Desirably higher than the softening temperature of the material.

Further, the porosity of the partition wall 4 is such that it has a strength that does not hinder the handling and is easy to carry the above-mentioned hydroxyl group.
0.05 to 25%, especially 0.1 to 20%, and even 0.1 to 0.2%.
It is desirably 15 to 10%.

The shape of the recess 2 is shown in FIGS.
As shown in the schematic perspective view of (c), (a) a plurality of rib-shaped partition walls 4 are arranged in parallel to form a groove-shaped recess 2, and (b) a plurality of roughly circular recesses 2. Side by side shape,
(C) A shape in which the grid-like partition walls 4 are formed and the concave portions 2 are formed between the grids, etc., can be surely divided between the spots, and the structure of (c) is employed in terms of productivity. Is desirable.

According to the present invention, the pitch interval W between the concave portions 2 is 500 μm or less, particularly 300 μm or less, and more preferably 20 μm or less.
It is important that the thickness is 0 μm or less, and one spot is dropped and stored in each of the recesses defined by the partition walls 4 so that the spots are not mixed with each other and a large amount of sample can be processed in a small space. Can be. Note that the pitch interval between the concave portions 2 in the present invention is shown in FIG.
As shown in FIG. 2, the distance W between the centers of the closest partition walls 4, 4 means the distance W.

Also, in the structure shown in FIG. 2A, each spot is held in a state of contact between the adjacent partitions 4, 4, so that the spot holding force is higher than in the conventional case where there is no partition and no concave portion. Increase.

Further, the height of the partition wall 4 is 30 μm or more,
In particular, 50 μm or more, further 100 μm or more, furthermore 2
It is desirable that the thickness be not less than 00 μm.

Here, for example, the shape of the partition wall 4 in the shape of (a) is rectangular or trapezoidal in cross section.
The wall surface is concave curved surface such as arc, ellipse, parabola etc.
Desirably, the thickness is 450 μm. Further, for example, in the shape of (b), it is desirable that the substantially circular concave portions 2 having a diameter of 50 to 490 μm, particularly about 100 to 300 μm are arranged in a line. Furthermore, for example, in the shape of (c), it is desirable that the partition walls 4 having a thickness of about 30 to 450 μm are arranged in a grid pattern.

The shape of the bottom of the recess 2 is desirably a curved surface in order to enhance the ease of handling and the mechanical reliability of the container 1.

Next, an example of the method for producing the sample container for biochemical analysis of the present invention will be described with reference to the process chart of FIG.
First, a known printing method such as screen printing is repeated a plurality of times using a paste for the partition walls 4 on the surface of the substrate 3 made of glass as described above. As shown in FIG. A mold 12 having a groove 11 for the partition 4 on the surface of the paste layer 10
After plastic deformation of the mold 0, a molded body 13 for the partition wall 4 is manufactured by a stamping method of releasing the mold 12 or a method of etching a portion other than the partition wall 4 by a photoresist method.

When the mold 12 is used, the width at the bottom of the groove 11 for the partition 4 of the mold 12 is smaller than the width at the opening of the groove 11 for the partition 4 of the mold, and particularly at the bottom of the groove. By forming the wall surface to have a concave curved surface, the releasability of the molded body 13 for the partition wall 4 from the molding die 12 is further improved. In addition, a roll-shaped mold may be used as the mold 12 in terms of mold release properties and productivity of the mold 12, and in order to manufacture a container having the above-described shape using this roll-shaped mold, A mold as shown in FIG. 4 may be used.

Then, the substrate 3 on which the molded body 13 for the partition wall 4 is adhered is formed, for example, in an oxidizing atmosphere at 550-650.
By baking at ℃, the sample container for biochemical analysis of the present invention can be prepared.

In order to produce the sample container for biochemical analysis of the present invention, in addition to the above-mentioned methods, a method of grinding the surface of a substrate made of glass or ceramics using a slicer, a drill or a sand blast method, or a laser. A method of forming a predetermined concave portion by irradiation can also be adopted.

[0027]

(Embodiment 1) 100 mm × 100 mm × 1.
Si, Al, Z having a strain point of 450 ° C. and an average particle size of 5 μm are formed on a glass substrate surface made of 3 mm soda lime glass.
Titanium, alumina, fused silica, and α-
Terpineol, polyvinyl butyral, and a dispersant,
A paste mixed with a solvent such as alcohol and kneaded was formed into a layer by a slit coater, and the paste layer was dried to a hardness that allows plastic deformation.

On the other hand, a mold having a 50.times.50.times.5 mm metal flat plate having a partition groove on the surface thereof is 100 .mu.m wide and 50 .mu.m long.
A flat plate having protrusions having a thickness of 100 μm and a depth of 100 μm was arranged at a pitch of 200 μm, and was pressed so that the protrusions of the flat plate were embedded in the surface of the paste layer, and the mold was released.

This was fired at 560 ° C. in the air to obtain a sample container having a groove-shaped concave portion having a width of 100 μm, a height of 100 μm, a length of 50 mm and a pitch of 200 μm on the substrate surface. The porosity of the partition wall was measured by the Archimedes method and found to be 0.2%.

Another flat plate is placed on the opening surface of the concave portion of the obtained sample container, a force for compressing the container is applied in the height direction of the container, and a load F at which the partition wall breaks is measured. Cross-sectional area S
When the pressure P (F / S) with respect to was calculated as the strength of the container, it was 0.1 MPa, suggesting that the container was durable.

Further, the amount of metal ions eluted after filling the obtained sample container with water and holding it for one hour was measured by ICP analysis. As a result, it was lower than 0.3 ppm.

Further, steam 12 which is a condition for sterilization treatment
The dimensional change rate after exposing the sample container to 0 ° C. and 0.4 MPa for 2 hours is 1 × 10 ⁻⁴ % or less, and the strength reduction rate is 0.1% or less.
It was also suggested that repeated use is possible.

(Example 2) The protrusions of the mold of Example 1 were formed such that the thickness of the frame was 100 μm, and the concave shape between the frames was 200 μm × 20.
A sample container was prepared and evaluated in the same manner as in Example 1 except that the sample container was formed in a grid shape of 0 μm, and as a result, the strength was 0.3 MPa.

[0034]

As described in detail above, according to the sample container for biochemical analysis of the present invention, by using a container having a plurality of recesses formed on the surface of a plate-like body made of glass and / or ceramics. In addition, a large amount of sample can be efficiently processed, sterilization treatment under high temperature and high pressure can be performed, and the sample container for biochemical analysis can be used repeatedly.

[Brief description of the drawings]

FIG. 1 is a schematic sectional view of a sample container for biochemical analysis of the present invention.

FIG. 2 is a schematic perspective view showing an example of a sample container for biochemical analysis of the present invention.

FIG. 3 is a process chart showing an example of the method for producing a sample container for biochemical analysis of the present invention.

FIG. 4 is a view for explaining an example of a molding die in the molding step of FIG. 3;

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Sample container for biochemical analysis 2 Concave part 4 Substrate 5 Partition wall

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Kiyohiro Sakasegawa 1-4-4 Yamashita-cho, Kokubu-shi, Kagoshima Inside the Kyocera Research Institute (72) Inventor Kazuo Watada 1166-6-1, Haseno, Hachimizo-cho, Yokaichi-shi, Shiga Prefecture. Kyocera Corporation Shiga Factory F-term (reference) 2G058 CC00 CC02 CC08 4B029 AA08 BB20 GA03 GB04 GB09 GB10

Claims (6)

[Claims] 1. A sample container for biochemical analysis comprising a plurality of recesses formed at a pitch of 500 μm or less on the surface of a substrate made of glass and / or ceramics. 2. The sample container for biochemical analysis according to claim 1, wherein said concave portion has a groove shape. 3. The sample container for biochemical analysis according to claim 1, wherein the partition walls between the concave portions are formed in a lattice shape. 4. The biochemical analysis sample container according to claim 1, wherein the amount of metal ions eluted after filling the concave portion with water for 1 hour is 1 ppm or less. . 5. The sample container for biochemical analysis according to claim 1, wherein the content of alkali metal in said glass and / or ceramic is 5% or less. 6. The sample container for biochemical analysis according to claim 1, which is used for DNA analysis.