JP2000093816A - Production of small-sized experimental plate and producing device for in-line type small-sized experimental plate - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a producing method or the like of a small-sized experimental plate (one tip experimental chamber) free from incorporating cut chips or foreign matter in a channel (groove). SOLUTION: The producing method of the small-sized experimental plate is provided with a 1st member (lower part plate 1 or the like) having at least the channel (groove) 3 and a 2nd member (upper part plate 2 or the like) having a part to be a liquid storage part 5 and has a structure obtained by integrating the 1st member and the 2nd member so that the channel 3 becomes a passage of liquid in the liquid storage part 5. In such a case, a process for forming the 1st member by preparing a portion of a material (preform or the like) for the 1st member necessary for forming one small-sized experimental plate and forming the channel 3 thereon, a process for forming the 2nd member by forming the part to be the liquid storage part 5 and a process for integrating the 1st member and the 2nd member are included.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a method for separating, reacting, purifying a liquid by freely moving a liquid by electrophoresis or the like on a plate (substrate) on which fine channels (grooves) are formed.
The present invention relates to a method of manufacturing a small experimental plate (so-called one-chip laboratory) for performing operations such as DNA division.

[0002]

2. Description of the Related Art The main laboratory equipment used in laboratories, such as bio-related (biological, biochemical, medicinal, life science, food, etc.) and chemical / environmental related labor, which previously occupied the desk, was enlarged. A small experimental plate integrated and integrated on a substrate just a few centimeters in size has recently been attracting attention by a US venture company. This small experimental plate uses microfabrication technology on the substrate surface to form micron-order grooves and holes, etc., and can freely configure experimental equipment by changing the layout design of grooves and holes. Because it resembles a chip, it is called a one-chip laboratory or a lab chip. Using such a small experimental plate (hereinafter referred to as a one-chip laboratory), various experiments that have been conducted by researchers in the laboratory using pipettes, test tubes, beakers, flasks, and the like,
On a chip of a few centimeters square, a very small amount of reagents enables rapid experimental work and analysis, and enables many systematic experiments to be performed efficiently and quickly. In such a one-chip laboratory, a voltage is applied to a part of the chip to move the sample in the groove or hole to cause a reaction, and all operations such as electrophoresis, protein analysis, and enzyme reaction are performed. Experiments can be performed on the chip, such as search for new drugs, drug development, gene analysis (including gene activity investigation and gene identification), DNA manipulation, nucleotide sequence analysis, pathological examination, gene diagnosis, etc. It is expected to be effective in speeding up and reducing costs. Identification of the generated product is often performed by optical measurement such as fluorescence analysis or absorption spectrum.

The one-chip laboratory is, for example, as shown in FIGS. 1 and 2, a lower plate 1 in which a channel (groove) 3 having a width and a depth of about several tens μm is engraved and bonded to the lower plate 1. The upper plate 2 forms an upper portion of the channel 3, and a through hole 4 communicating with the channel 3 is formed in a part of the upper plate 2 to serve as a liquid storage unit 5 (liquid reservoir, sample supply unit). Fulfill. And, for example,
An electrode is set in the storage part, a potential difference is applied, and the liquid in the storage part or the channel is freely moved by electrophoresis, and operations such as separation, reaction, purification, and DNA division are performed. The electrodes are usually inserted externally. The reaction time can also be changed by changing the mixing ratio of the two kinds of liquids or changing the applied voltage to change the speed of electrophoresis.

[0004]

As a method of forming a channel in a lower plate, an etching method by photolithography is known, and a plurality of chips are collectively formed on one large plate-like material like a semiconductor chip. Processing to increase productivity. In this method, after the channel is processed, the plate on which the channel is formed must be cut off one by one according to the size of the chip. At this time, cutting chips may enter the channel. As described above, since the channels are minute, it is not easy to remove the cutting waste once entered. In order to avoid this, it is conceivable to cut the channel after completely covering the channel with resin or the like, and then remove the resin after cutting. However, since the resin must not be left in the channel, there is a problem that the process is troublesome and it takes time to check whether the resin is completely removed. After forming channels for multiple chips in the large lower plate, it may be integrated with the large upper plate with holes for the liquid storage part for multiple chips, and then cut into individual chips. Therefore, it is very difficult to remove the cutting debris once mixed in the channel because the channel does not protrude on the surface. Therefore, there is a problem that protection against cutting-off chips from entering the channel needs to be performed more carefully.

The present invention has been made in view of the above background, and has as its object to provide a method of manufacturing a small experimental plate (one-chip laboratory) in which cutting chips and foreign matter are not mixed into a channel or the like. I do.

[0006]

In order to achieve the above object, a method of manufacturing a small-sized experimental plate (one-chip laboratory) according to the present invention has the following configuration.

(Structure 1) At least a channel (groove)
And a second member (e.g., an upper plate) having a portion serving as a liquid storage portion, such that the channel serves as a passage for the liquid in the liquid storage portion. A method for manufacturing a small test plate having a structure in which the first member and the second member are integrated, wherein a material of the first member required for forming one small test plate ( A step of preparing a first member by forming the channel therein, a step of forming a portion to be the liquid storage portion and forming a second member, And a step of integrating the second member with the second member.

(Structure 2) The method according to Structure 1, wherein the first member is manufactured by forming a channel by molding.

(Structure 3) The small-sized experiment according to Structure 2, wherein the first member and the second member are integrated by thermocompression bonding, following the molding process of the first member. Plate manufacturing method.

(Structure 4) The structure according to Structure 2 or 3, wherein the molding of the first member is performed by using a mold in which at least a portion for transferring and molding the channel is formed by photolithography. Manufacturing method of small experimental plate.

(Structure 5) The method according to any one of Structures 1 to 4, wherein the first member and the second member are made of glass.

(Structure 6) A means for supplying a preform to be used for the lower plate, a mold used for molding the preform, a mold driving means for performing molding and releasing, Means for transporting the lower plate in the apparatus, means for supplying the upper plate onto the molded lower plate, means for performing thermocompression bonding between the lower plate and the upper plate, and small-sized thermocompression bonding. An apparatus for manufacturing an inline-type small experimental plate, comprising: means for collecting an experimental plate; and means for controlling a temperature in each step.

[0013]

According to the above configuration 1, when producing the first member (the lower plate or the like), the material of the first member necessary for forming one small experimental plate is prepared,
A channel is formed in this to produce a first member. In this case, prior to the channel formation, the material of the first member is already in a size (amount) corresponding to one small experimental plate.
Therefore, after forming the channel, the first member is
There is no need to cut to a size corresponding to an individual. Therefore, cutting chips and foreign matter do not enter the channel.

According to the above configuration 2, by using the molding method for forming the first member (such as the lower plate),
No debris is generated during channel processing, and the channel processing can be performed with extremely high reproducibility. In addition, as a material of the first member used in the molding method, a preform, a glass gob, a molten glass, or the like can be used.

According to the above configuration 3, by continuously performing the molding and the thermocompression bonding, even if microorganisms adhere to the channel or the like, the same effect as the heat sterilization can be obtained, and the channel or the like is biochemically contaminated. Is extremely low.

According to the above configuration 4, the first member is molded by using a mold in which at least a portion for transferring and molding the channel is formed by using the photolithography technique, so that the reproducibility is extremely high. Channels and the like can be formed with high precision.

According to the above configuration 5, it is preferable that the first member and the second member are formed of glass in terms of chemical durability and optical transparency.

According to the apparatus having the above configuration 6, a small experimental plate can be manufactured efficiently.

[0019]

Embodiments of the present invention will be described below.

Small experimental plate (one-chip laboratory)
Comprises a first member and a second member, the first member is provided with a minute channel for moving liquid, and the second member communicates with the channel and moves the channel. A portion serving as a liquid storage portion for storing the liquid to be formed is formed.

Here, both the first member and the second member are not particularly limited in terms of their shapes and the positional relationship between the members, and can be freely designed according to the purpose. For example, the shape may be a substrate capable of forming a channel, and may be a flat plate (circle, rectangle, square, etc.), a three-dimensional structure, or a shape other than the flat plate (eg, block shape).

Here, a case where both the first member and the second member have a plate shape and the second member is located above the first member in an integrated state will be described as an example. Here, the first member is called a lower plate, and the second member is called an upper plate.

In manufacturing the lower plate, a material for the lower plate used for one chip in the one-chip laboratory is prepared, and a channel is formed in the material. Prior to forming the channel, the material of the lower plate is already prepared in a necessary amount for one chip, so that it is not necessary to cut the lower plate into a single size after the channel is formed. Therefore, cutting chips and foreign matter do not enter the channel.

For the formation of the lower plate, a method that does not generate debris during channel processing, such as a molding method or a processing method by photolithography, can be used. In particular, molding is preferable because channel processing can be performed with very high reproducibility. A processing method using a photolithography method (such as etching) is inferior to a molding method in terms of mass productivity, but does not require the formation of a mold, and is effective when a prototype chip of a new design is manufactured.

The material of the lower plate is not particularly limited, and examples thereof include glass, plastic, silicon, ceramic, and glass ceramic. Among these materials, glass, plastic, etc. are preferable from the viewpoint of a material that has electrical insulation properties so that a liquid can be freely manipulated by an electrophoresis method and is easy to mold, but is chemically durable or optically durable. Glass is particularly preferable from the viewpoint of a material having a high degree of transparency (for reasons such as the convenience of performing optical measurements) and from the viewpoint of the results of use of laboratory equipment made of glass.
The lower plate material and glass material are not only materials that are easy to mold, but also have resistance to weak acids and weak alkalis, as well as wettability, water repellency, surface tension, elution of glass components, glass surface polarity and surface It is preferable to select in consideration of factors that affect the sample such as a group. Many optical glasses can be used as the glass material, but it is better to avoid materials containing lead or fluorine in order to prevent fusion with a mold at the time of molding. Examples of the glass material include borosilicate based materials such as white plate (BK7), and La
Many glasses such as glass, Zr, and Ti can be used.

When manufacturing the lower plate by molding, it is necessary to mold a fine portion such as a channel with high precision. Therefore, it is necessary to use a mold in which at least a portion for transferring and molding the channel (mold) is formed by photolithography. Is desirable. As the mold material, quartz, silicon carbide, or the like can be used. If desired, a mold release film may be formed on the molding surface via an intermediate film or directly, in order to enhance mold release properties. By using such a mold, channels and the like can be formed with very high reproducibility and high accuracy, and the cross-sectional shape and the like of the channels can be uniform over the entire surface or as designed. When molding the lower plate, a normal precision molding technique can be used.

The channel size (depth, width), cross-sectional shape, gradient as a water channel (gradient for facilitating flow from a high place to a low place), layout, and the like can be appropriately designed according to the use and purpose. For example, the cross-sectional shape can be a rectangle, an inverted trapezoid, an inverted triangle, a U-shape, a semicircle, or the like. Further, the depth, width, gradient, cross-sectional shape and the like of the channel can be partially changed, and these can also be changed in the length direction of the channel. In addition, the factors that affect the sample, such as durability, wettability, water repellency, surface tension, elution of glass components, polarity of the glass surface, and surface groups, are entirely or partially improved in the channel or the liquid storage part. Therefore, various surface treatments can be performed.

In the processing of the liquid storage portion of the upper plate, since the liquid storage portion is not minute like a channel, even if cutting debris generated during processing adheres, it is easily removed by washing or the like. can do. Therefore, processing of the upper plate does not require special consideration as in the processing of the lower plate, and emphasizes productivity and uses general processing such as ultrasonic processing, water jet processing, and laser processing on large plate-like materials. It is also possible to use a method of forming a part to be a liquid storage part by making a hole by a suitable method, and then cutting it to the size used for one chip, or adopting molding as in the case of the lower plate it can.
Like the lower plate, glass is preferably used as the material of the upper plate from the viewpoint of a material having electrical insulation, chemical durability, and optical transparency. In addition,
The lower plate and the upper plate can be formed of different materials, for example, the lower plate can be formed of glass and the upper plate can be formed of plastic, and the upper and lower plates can be formed of different glass materials.

The lower plate and the upper plate manufactured as described above are aligned and integrated. Prior to the integration, if there is a possibility that cutting chips generated during processing may adhere to the upper plate, cleaning and the like are performed to remove cutting chips. The method of integrating the lower plate and the upper plate is not particularly limited, and a known bonding method such as thermocompression bonding or adhesion can be used, but thermocompression bonding is preferable. Particularly preferred is a method in which the lower plate material is hot-molded, and the lower plate and the upper plate are integrated by thermocompression bonding continuously with the molding. By performing thermocompression bonding, the possibility of biochemical contamination of the channel is greatly reduced. For example, even if microorganisms adhere to the channel, the same effect as heat sterilization can be obtained by continuously performing thermocompression bonding or molding and thermocompression bonding. In addition, if the lower plate and the lower plate and the upper plate are integrated by thermocompression bonding continuously, after molding,
There is no need to lower the temperature of the lower plate to room temperature, and efficient and high productivity can be obtained.

One embodiment of the method and apparatus of the present invention will be described in detail with reference to FIG. On the left side of FIG. 3 (a), a glass preform used for the lower plate, a mold used for molding the preform, and a portion serving as a liquid storage portion are formed and the upper portion is cleaned. The plate is being prepared. The glass preform used for the lower plate is, for example,
It is formed by cutting and polishing a glass substrate or a glass ingot, or by molding a molten glass. Similarly, the upper plate is formed by, for example, cutting, polishing, and perforating a glass substrate or a glass ingot. A portion surrounded by a broken line in FIG. 3A indicates an in-line press device, and the mold 12 is attached to this device. FIG. 3B shows an outline of a temperature change of the lower plate and the like in each step of the in-line press device. First, the glass preform 11 for the lower plate is loaded into the apparatus (zone 1), and the temperature is raised to the temperature at which molding is performed (zone 2). When the molding temperature is reached, molding is performed (zone 3), and then the temperature is lowered to an intermediate temperature for release (zone 4). After the temperature is lowered to the intermediate temperature, the mold is released and the mold 12 is collected (zone 5).
The recovered mold 12 is used for molding a new lower plate. Next, the upper plate 13 is loaded into the apparatus (zone 6), and the temperature is raised to the thermocompression bonding temperature together with the lower plate (zone 7). When the temperature reaches the thermocompression bonding temperature, the lower plate and the upper plate are thermocompression bonded (zone 8), the temperature is reduced to a temperature at which the lower plate can be taken out (zone 9), and the chips 14 are collected (zone 10). FIG.
The outline of the temperature change of the lower plate (including from the stage of the preform to the chip integrated with the upper plate to the chip) in zones of 10 to 10 is shown. Since the glass loaded into the mold pressing device is placed under high temperature up to the zone 10, a finished chip can be obtained without biochemical contamination of the formed channel. Also, since the molded lower plate is not cooled to room temperature, reheating of the lower plate to the thermocompression bonding temperature can be minimized. As described above, the lower plate and the upper plate are integrated to obtain a one-chip laboratory without mixing chips or the like in the channel and without taking any special measures to protect the mixing of chips. be able to. In addition, the whole device or the carrying-out part of the device can be covered with a sterile booth, and the carried-out chips can be aseptically packaged.

Embodiments of the present invention other than those described above will be described below. The method of the present invention can also be used when there are a plurality of members corresponding to the first member and / or a plurality of members corresponding to the second member. In addition, a channel can be formed in the second member. In this case, the second member can be manufactured in the same manner as the first member. Specifically, for example, a portion serving as a liquid storage portion is processed in advance in a material used for one chip, and then a channel is formed, or a portion serving as a liquid storage portion and a channel are formed. Are collectively molded and formed. In these cases, no cutting waste is generated after the channel is formed. Further, the first member and / or the second member can be provided with an electrode, a wiring, a biosensor, a detection sensor or a detection element, and a DNA fragment, a reagent, a solvent, and the like are provided. Can be. The method of the present invention can also be used when only the first member is used as a one-chip laboratory. In this case, the liquid can be directly supplied from the outside to the channel of the first member and used, or the channel and the means for supplying the liquid to the channel (for example, a liquid storage portion) are formed in the first member. Can also be used. Further, a first member and a second member or a third member may be added to the first member and the second member, and a laminated structure or a three-dimensional structure such as a semiconductor element may be formed using these members. Further, the size of the first member and the size of the second member may not be the same. For example, the second member may be partially joined on the first member.

[0032]

An embodiment of the present invention will be described below with reference to FIG.

Plate-shaped quartz is used as a mold material for molding the lower plate, which is the first member, and the surface of the quartz plate is subjected to photolithography (resist spin coating, exposure, development, dry etching (RIE)). ) To form a projection for transfer-molding a channel having a width of 35 μm and a height of 10 μm. A 100 nm thick SiC sputtered film is provided as an intermediate film on the surface of the surface on which the convex portions are formed,
An i-carbo having a thickness of 100 nm is formed thereon as a release film.
The n-film was coated to form a mold for molding the lower plate (see step A in FIG. 4).

Using this mold, a 20 mm square, 1 mm thick borosilicate glass (glass transition temperature: 480 ° C.) molding material was molded to obtain a lower plate having channels. The molding conditions are as follows: molding temperature 55
The temperature was 0 ° C., the molding load was 50 kg, and the holding time was 3 minutes (see step B in FIG. 4).

On the other hand, the same glass as the lower plate is used.
A material having a size of 1 mm square and a thickness of 1 mm was prepared, and a through hole for a liquid storage section having a diameter of 2 mm was formed at a predetermined location by ultrasonic processing to form an upper plate. A plurality of upper plates may be manufactured by forming through holes for a plurality of upper plates in a large glass plate and then cutting to a predetermined size (see step C in FIG. 4). . The upper plate manufactured in this manner was washed to remove processing and cutting debris attached during processing.

Next, the lower plate was positioned so that the channel of the lower plate communicated with the through-hole for the liquid storage section of the upper plate, and the lower plate and the upper plate were integrated by thermocompression bonding. The conditions of thermocompression bonding are as follows: temperature 520 ° C, load 50k
g and the retention time was 2 minutes. Thus, the lower plate and the upper plate were integrated to obtain a one-chip laboratory chip (see step D in FIG. 4).

When the channels of the manufactured chip were observed with an optical microscope and an electron microscope, no processing / cut debris or foreign matter was found (see FIG. 5).

[0038]

As described above, according to the method for manufacturing a small experimental plate of the present invention, a small experimental plate (one-chip laboratory) can be manufactured without mixing processing / cutting debris or foreign matter into the channel. Can be. Therefore, conventionally, the debris which cannot be removed by any means mixed with the channel is regarded as a defective product, but the production method of the present invention improves the yield. Further, since it is not necessary to take any special measures for protecting the channel from dust and foreign matter, the manufacturing process is simplified and the productivity is improved. Further, according to the apparatus for manufacturing a small experimental plate of the present invention, a small experimental plate can be efficiently manufactured.

[Brief description of the drawings]

FIG. 1 is a partial cross-sectional view illustrating a small experimental plate.

FIG. 2 is a plan view for explaining a small experimental plate.

3A and 3B are diagrams illustrating a method and an apparatus according to one embodiment of the present invention, and FIG. 3A is a diagram illustrating an outline of an in-line press device and the like, and FIG. Figure 3
It is a figure for explaining an outline of a temperature change in each zone in (a).

FIG. 4 is a diagram schematically showing a flow of a manufacturing process of a small experimental plate in an example.

FIG. 5 is an electron micrograph of the vicinity of a channel of a small experimental plate manufactured in an example.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Lower plate 2 Upper plate 3 Channel (groove) 4 Through-hole 5 Liquid storage part 11 Glass preform 12 type 13 Upper plate 14 Chip

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12M 1/00 G01N 27/26 331E

Claims (6)

[Claims] At least a first member having a channel (groove) and a second member having a portion serving as a liquid storage portion, wherein the channel serves as a passage for a liquid in the liquid storage portion. A method for manufacturing a small test plate having a structure in which the first member and the second member are integrated, the material of the first member being necessary to form one small test plate; Preparing a first member by forming the channel therein, a step of forming a portion to be the liquid storage portion to prepare a second member, and the first member and the second member And a step of integrating the two members with each other. 2. The method according to claim 1, wherein the first member is manufactured by forming a channel by molding. 3. The small experimental plate according to claim 2, wherein the first member and the second member are integrated by thermocompression bonding, following the step of molding the first member. Manufacturing method. 4. The compact device according to claim 2, wherein the first member is molded using a mold in which at least a portion for transferring and molding the channel is formed by photolithography. How to make an experimental plate. 5. The method according to claim 1, wherein the first member and the second member are made of glass. 6. A means for supplying a preform to be used for the lower plate, a mold used for molding the preform, a mold driving means for performing molding and release, and a molded lower part Means for transporting the plate in the apparatus, means for supplying the upper plate on the molded lower plate, means for performing thermocompression bonding between the lower plate and the upper plate, and a small experimental plate obtained by thermocompression bonding And a means for controlling a temperature in each step.