JP2003254875A - Treatment method for specimen and tip set - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a treatment method for a specimen in which the specimen in a small amount can be treated and in which an object (an object component) can be sampled easily and surely from the specimen and to provide a tip set used for the treatment method for the specimen. <P>SOLUTION: In the treatment method for the specimen, the specimen and a reagent containing particles capable of carrying the object are sucked into a tip T mounted on a tip part of a nozzle 30 at a dispensing device 100, and the object in the specimen is carried by the particles. In a state that a filter- attached member 2 which is provided with a tubular member 21 mountable on the tip part of the tip 1 and a filter 22 installed in a flow channel of the tubular member 21 so as to be capable of capturing the particles is mounted on the tip part of the tip 1, a mixed solution of the specimen and the reagent is discharged from the tip 1, and the particles carrying the object are captured by the filter 22. It is preferable to perform an operation to stir the mixed solution of the specimen and the reagent after the specimen and the reagent have been sucked into the tip 1. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sample processing method and a chip set.

[0002]

2. Description of the Related Art Conventionally, in a wide range of fields such as medical care, pharmaceuticals, living things, and foods, work (collection) of a target substance (target component) from a sample such as blood for the purpose of inspection, measurement, and research is necessary.

For example, in the enzyme immunoassay (EIA method), particles coated with an antigen or antibody are
In the ELISA method, a target substance is reacted (for example, adsorbed) on a plate in which the surface of a microplate is coated with an antibody or the like, and then collected.

Next, an operation (B / F separation process) of removing unnecessary substances other than the target substance is performed by washing the particles and the microplate with a large amount of cleaning liquid. Such an operation is usually performed using an automated device.

However, this apparatus requires a dispensing means for supplying (dispensing) a reagent, a sample, a cleaning liquid, etc. to a reaction container, and a discharging means for discharging the cleaning liquid. Therefore, there is a problem that the device becomes large in size and the cost becomes high.

Further, in the method using particles, in order to avoid erroneous discharge of particles in the washing step, it is necessary to control the position of the discharging means with high accuracy, and the size (particle size) of usable particles is limited. There was a problem that there is.

On the other hand, in the method using the microplate, it is necessary to control the position of the discharging means with high accuracy so that the coating on the surface is not peeled off.
In the washing step, there is a problem that the removal rate of unnecessary substances (unnecessary components) other than the target substance varies, and the purity of the obtained target substance decreases (the amount of the unwanted substance mixed increases).

Further, a reaction vessel (in the method using particles,
For example, in the method using a test tube or a microplate, since the volume of each well of the microplate) is large, there is a problem that a large amount of reagents, specimens, washing solutions, etc. are required, resulting in high cost.

[0009]

SUMMARY OF THE INVENTION An object of the present invention is to process a small amount of a sample, and to collect a target substance from the sample easily and surely, and a method for processing a sample, It is to provide a chip set used in such a sample processing method.

[0010]

Such an object is achieved by the present invention described in (1) to (22) below.

(1) A sample and a reagent containing particles capable of supporting a target substance are sucked into a tip attached to the tip of a nozzle, and then a tubular member attachable to the tip of the tip, A filter-equipped member provided in the flow path of the tubular member and provided with a filter capable of capturing the particles is attached to the tip of the chip, and a mixed liquid of the sample and the reagent is discharged from the inside of the chip. By doing so, the method of treating a specimen is characterized in that the particles carrying the target substance are captured by the filter.

(2) The method for treating a sample according to the above (1), wherein after the sample and the reagent are sucked into the chip, an operation of stirring the mixed solution is performed.

(3) The stirring operation is performed by discharging the mixed solution from the inside of the chip and then again sucking into the inside of the chip at least once. Processing method.

(4) The method for treating a sample according to any one of the above (1) to (3), wherein the particles are loaded with the target substance while heating a mixed liquid of the sample and the reagent.

(5) The heating temperature is 20 to 100.
The method for treating a specimen according to (4) above, which is at a temperature of 0 ° C.

(6) The method for treating a specimen according to any one of the above (1) to (5), wherein the target substance is supported on the particles while irradiating a mixed liquid of the specimen and the reagent with electromagnetic waves.

(7) The method for treating a sample according to (6), wherein the electromagnetic wave is a microwave.

(8) The method for treating a sample according to (6), wherein the electromagnetic wave is ultraviolet light.

(9) The method for treating a specimen according to any one of the above (1) to (8), wherein at least the surface of the particles is solid-phased with a substance capable of supporting the target substance.

(10) The average particle size of the particles is from 10 to 10.
The method for treating a specimen according to any one of (1) to (9) above, which has a thickness of 1000 μm.

(11) After the particles carrying the target substance are captured by the filter, the washing liquid is sucked into the member with the filter, and then the washing liquid is discharged at least once, thereby The method for treating a sample according to any one of (1) to (10) above, wherein the particles supporting the target substance are washed.

(12) After the particles carrying the target substance are captured by the filter, a separation liquid for separating the target substance from the particles is sucked into the member with the filter, and then the separation liquid is removed. The method for treating a specimen according to any one of (1) to (11) above, wherein the target substance separated from the particles is recovered in the separated liquid by performing the discharging operation at least once.

(13) The method for treating a specimen according to any one of (1) to (12) above, wherein the filter has a property of not substantially adsorbing the target substance.

(14) The method for treating a specimen according to any one of (1) to (13), wherein the tubular member has a shape substantially equal to the shape of the tip.

(15) The method for treating a specimen according to any one of the above (1) to (13), wherein the tip has a reduced diameter portion in which the outer diameter and the inner diameter are reduced substantially at the center in the longitudinal direction.

(16) The method for treating a specimen according to (15), wherein the inner diameter and the outer diameter of the reduced diameter portion are substantially constant.

(17) The inner diameter (average) of the reduced diameter portion is
The method for treating a specimen according to (15) or (16), which has a thickness of 0.1 to 2 mm.

(18) The capacity of the reduced diameter portion is 0.1 to
The method for treating a specimen according to any one of (15) to (17) above, which is 100 μL.

(19) The sample processing method according to any one of (1) to (18) above, wherein the chip and the filter-equipped member are disposable after each sample processing.

(20) The object is a microorganism, nucleic acid,
The method for treating a specimen according to any one of (1) to (19) above, which is a protein, hormone, amino acid, peptide or nucleotide.

(21) By operating a pipetting device equipped with a pipetting pump for sucking and dispensing a liquid, the sample is sucked and transferred into the chip to process the sample. 20. The method for treating a sample according to any one of 20).

(22) A chip set used in the method for treating a sample according to any one of (1) to (21), wherein the chip is attachable to the tip of the nozzle of the dispensing device, and A chip set comprising: a tubular member attachable to a tip portion and a filter-equipped member having a filter provided in a flow path of the tubular member.

[0033]

BEST MODE FOR CARRYING OUT THE INVENTION The sample processing method and chip set of the present invention will be described below in detail with reference to the preferred embodiments shown in the accompanying drawings.

1 to 4 are schematic views showing the steps of the embodiment of the sample processing method of the present invention, respectively.
FIG. 6 is a partial cross-sectional view showing the configuration of the chipset of the present invention and a dispensing device used in the present invention, and FIG. 6 is a partial cross-sectional view showing another configuration example of the chip. In addition, in the following description, in FIGS. 1 to 6, the upper side is referred to as a “base end” and the lower side is referred to as a “tip”.

In the method for treating a sample shown in FIGS. 1 to 4, particles P containing a target substance (target component) Q in the sample S in a reagent R are used.
Carrying step [S1] for carrying the target Q, a collecting step [S2] for collecting the particles P carrying the target Q, a washing step [S3] for cleaning the particles carrying the target Q, and the target Q And a collecting step [S4].

Such a sample processing method is performed by using, for example, a dispensing apparatus 10 as shown in FIG.

The dispensing device 10 is composed of, for example, a piston type pump, a syringe type (plunger type) pump, etc., and is provided at a lower end portion (tip portion) of a dispensing pump 20 for sucking and dispensing a liquid. And a nozzle 30 which is provided.
Further, the dispensing device 10 can be moved in a three-dimensional direction by a moving mechanism (described later).

The nozzle 30 has a substantially cylindrical shape and is arranged so that its longitudinal direction is the vertical direction in FIG.
A passage 31 is formed inside the nozzle 30. The base end of the passage 31 communicates with the pump space (not shown) of the dispensing pump 20, and the air flows in and out of the pump space through the passage 31. On the other hand, the tip of the passage 31 is open to the tip of the nozzle 30.

The outer diameter of the nozzle 30 has a taper shape that gradually decreases toward the tip end. By inserting the tip end of the nozzle 30 into the base end opening of the chip 1 to be described later, The tip 1 can be easily and airtightly mounted on the tip of the nozzle 30.

In such a dispensing device 10, the volume of the pump space of the dispensing pump 20 is increased / decreased so that the liquid is sucked into the tip 1 and discharged from the tip 1.

In this embodiment, the sample S is sucked and transferred into the chip 1 by the operation of the dispensing device 10, and the following steps (processing of the sample S) are performed. Hereinafter, each step will be sequentially described.

[S1] Supporting Step (Supporting Operation) First, as shown in FIG. 5, the tip 1 is mounted on the tip of the nozzle 30 of the dispensing apparatus 10. This chip 1
It has an elongated tubular shape, and its outer diameter and inner diameter are gradually reduced toward the distal end (tapered).

Next, the sample S and the reagent R containing the particles P are sucked (introduced) into the chip 1 attached to the tip of the nozzle 30. At this time, the sample S and the reagent R may be sequentially sucked from different containers, or once, for example, the reagent R is supplied into the container in which the sample S is stored and the mixed liquid ( S + R), and the mixed liquid may be sucked. At this time, the target object Q in the sample S on the particle P
Are partially carried.

Next, the operation of stirring the mixed liquid (S + R) of the sample S and the reagent R that has been sucked (introduced) into the chip 1 is performed. As a result, the sample S and the reagent R can be mixed more uniformly, so that the particles P are contained in a larger amount of the target substance Q.
Can be efficiently carried.

Although any stirring method may be used, this stirring operation is performed by discharging from the inside of the chip 1 into the container (processing tube) X and then again sucking from the inside of the container X into the inside of the chip 1. Is preferred. According to this method, the sample S and the reagent R can be easily and reliably mixed uniformly.

It should be noted that such a stirring operation may be carried out only once, or may be repeated a plurality of times if necessary. That is, the stirring operation may be performed at least once. By repeating the stirring operation, the sample S and the reagent R can be mixed more uniformly, so that the particles P can more reliably carry the target substance Q. Further, this stirring operation can be omitted if necessary (for example, depending on the type of the target object Q, the ability of the particles P to support the target object Q, etc.).

The target Q to be collected according to the present invention is not particularly limited, but examples thereof include microorganisms such as viruses and bacteria, nucleic acids such as DNA and RNA, antigens, antibodies, and markers used for diagnosis of various diseases. And various hormones, amino acids, peptides, nucleotides and the like.

The sample S is not particularly limited, and can be, for example, a liquid in which blood, saliva, urine, biological tissue, agarose gel or the like is dissolved.

On the other hand, as the particles P, those in which at least the surface thereof is solid-phased with a substance capable of supporting the object Q are preferably used. As such particles P, for example, a surface of a base material made of a resin material or the like is solid-phased (coated) with a substance capable of supporting the target Q, and the whole is a substance capable of supporting the target Q. And the like.

As a substance capable of supporting the target substance Q, for example, groups having various charges (eg, sulfone group, carboxyl group, trimethylammoniomethyl group) are introduced to agarose, cellulose, nitrocellulose and the like. Examples thereof include ion exchange resins, various glasses, ceramic materials such as alumina and calcium phosphate, silica compounds such as amorphous silica and alkyl silica, resin materials such as polystyrene, polyethylene and polypropylene.

Depending on the type of the object Q, the following particles P can be used, for example. That is, when the target substance Q is a nucleic acid, the particles P may be those in which a nucleic acid having a structure complementary to the nucleic acid is immobilized on the surface of the base material. When the target substance Q is an antigen, the particles P may be those in which an antibody capable of specifically binding to the antigen is immobilized on the surface of a base material. When the target substance Q is an antibody, the particles P may be those in which an antigen capable of specifically binding to the antibody is immobilized on the surface of a base material.

The average particle size of the particles P is not particularly limited, but is preferably about 10 to 1000 μm.
It is more preferably about 0 to 500 μm, and 50 to 2
More preferably, it is about 00 μm. By setting the average particle diameter of the particles P in the above range, the particles P can secure a sufficient specific surface area and can efficiently carry the target Q.

Examples of the liquid component of the reagent R, that is, the dispersion liquid in which the particles P are dispersed (suspended) include various buffers (buffer solutions) such as a phosphate buffer, physiological saline and the like. . When the sample S is, for example, a biological tissue and the target substance Q is present in the sample S, the target substance Q is, for example, a protein denaturant from the sample S in the dispersion liquid. Efficient removal (elution)
You may make it add the substance which can be.

It is preferable that the target Q is supported by the particles P while heating the mixed solution. The heating of the mixed liquid can be performed by heating either the chip 1 or the container X, or by heating both of them. As a result, the particles P can efficiently carry the target Q.

The heating temperature is not particularly limited depending on the kind of the target product Q and the like, but is preferably about 20 to 100 ° C, more preferably about 37 to 95 ° C. If the heating temperature is too low, the particles P may not be able to efficiently carry the target Q depending on the type of the target Q, while if the heating temperature is too high, Depending on the type and the like, the target object Q may be altered or deteriorated.

Further, it is preferable to carry the target substance Q by the particles P while irradiating the mixed liquid with electromagnetic waves. Examples of this electromagnetic wave include microwaves and ultraviolet rays. In addition, these can also be used together.

By using the microwave, the efficiency of supporting the object Q by the particles P can be further increased. Further, by using ultraviolet rays, when the target Q is a microorganism such as a virus, it can be inactivated.

As described above, since the operation (supporting operation) for supporting the target substance Q on the particles P is carried out by using the mixed liquid (S + R) in an amount which can be stored in the chip 1, a small amount of the sample can be processed. It will be possible.

From the viewpoint of processing a small amount of sample, the capacity of the chip 1 is preferably about 50 to 500 μL, and more preferably about 100 to 300 μL.

[S2] Fractionation step (fractionation operation) Next, the filter-equipped member 2 is attached to the tip of the chip 1.

As shown in FIG. 5, the member 2 with a filter has an elongated tubular member 21 and a filter 22 provided so as to divide (separate) the inner cavity of the tubular member 21. By inserting and fitting the tip portion of the tip 1 into the base end opening of the tubular member 21, the member with filter 2 can be easily and airtightly attached to the tip portion of the tip 1.

The installation position of the filter 22 is not limited to that shown in the figure, and may be any position as long as it is installed in the flow path of the tubular member 21. That is, the filter 22 is the tubular member 2
It can be installed at any position in the up-down direction of the inner cavity of No. 1 or at the tip position of the tubular member 21 so as to close the tip opening.

In this embodiment, the tubular member 21 has a shape substantially equal to the shape of the tip 1. In other words, the tubular member 21 can be configured using the tip 1. In this case, there is an advantage that the manufacturing cost of the member with filter 2 can be reduced.

The constituent materials of the tubular member 21 and the tip 1 are, for example, polyvinyl chloride,
Polyethylene, polypropylene, polystyrene, poly-
Examples include various resins such as (4-methylpentene-1), polycarbonate, acrylic resin, acrylonitrile-butadiene-styrene copolymer, polyester such as polyethylene terephthalate, butadiene-styrene copolymer, and polyamide.

On the other hand, the filter 22 can capture the particles P. The pore size of the pores 221 of the filter 22 is set so that the particles P do not substantially pass through (that is, most of the particles P can be captured by the filter 22). As a result, the filter 22 keeps the target Q
It is possible to capture the particles P carrying the.

Further, it is preferable that the filter 22 has a property of not substantially adsorbing the target substance Q (that is, hardly adsorbing the target substance Q, or even adsorbing the target substance Q very little). Thereby, in the step [S4] described later, when the operation of recovering the target Q is performed, the filter 2
It is possible to prevent the target product Q from being adsorbed on the product 2 and reduce the recovery amount (yield) of the target product Q.

Such a filter 22 is obtained by, for example, being made of a material that does not easily adsorb the target Q, or by subjecting the surface of the base material to a surface treatment that makes it difficult to adsorb the target Q. be able to.

Examples of the material include polyamide (for example, nylon 6, nylon 6.6, nylon 6.
10, nylon 12), polypropylene, synthetic resins such as tetrafluoroethylene (PTFE), glass fibers, and the like, and one or more of these may be used in combination.

Examples of the surface treatment method include a method of coating the surface of the substrate with a silicone resin, tetrafluoroethylene or the like.

The filter 22 may have a property similar to that of the particles P, that is, a property capable of adsorbing the target substance Q. In this case, there is an advantage that it is possible to collect the target Q that cannot be fully supported by the particles P.

The form of the filter 22 is not particularly limited, and may be, for example, a woven fabric, a non-woven fabric, a foam (a sponge-like porous body having communication holes), or the like.

In this embodiment, the filter-equipped member 2 and the chip 1 constitute the chip set of the present invention.

Then, with the filter-equipped member 2 attached to the tip of the chip 1, an operation (preparation operation) for discharging the mixed solution (S + R) from the inside of the chip 1 is performed. As a result, the particles P carrying the target Q are captured by the filter 22.

At this time, the particles P carrying the target Q are
Although it cannot pass through the filter 22, unnecessary components (unnecessary components) O other than the liquid component and the target substance Q in the mixed liquid
Passes through the filter 22, is discharged to the outside of the member with filter 2, and is discarded.

[S3] Washing Step (Washing Operation) Next, the washing liquid W is sucked (introduced) into the filter member 22, and then the washing liquid W is discharged (washing operation). Thus, the particles P carrying the target Q are washed.

At this time, the particles P carrying the object Q are
The particles P and unnecessary substances (unnecessary components) O non-specifically attached to the filter-attached member 22 that cannot pass through the filter 22 are discharged to the outside of the filter-attached member 22 together with the cleaning liquid W.

By carrying out such a washing operation, it is possible to prevent or suppress the mixing of the unnecessary substance (unnecessary component) O into the obtained target substance Q.

The washing liquid W is not particularly limited, but examples thereof include distilled water, ion-exchanged water, RO water, various water such as pure water and ultrapure water, various alcohols such as ethanol, and phosphate buffer. Various buffers (buffer solutions) and the like.

Incidentally, such a washing operation may be carried out only once, or may be repeated a plurality of times, if necessary. That is, the cleaning operation may be performed at least once. By repeating the washing operation,
The removal rate of unnecessary substances (unnecessary components) O can be improved.

When the cleaning operation is repeated, the cleaning liquid W is replaced by a new cleaning liquid W for each cleaning operation.
It is preferable to replace the above with the above. As a result, the unnecessary substance (unnecessary component) O can be removed more reliably.

[S4] Recovery Step (Recovery Operation) Next, an operation (recovery operation) of sucking (introducing) the separated liquid T into the filter member 22 and then discharging the separated liquid T is performed. As a result, the target product Q separated from the particles P
Are collected in the separated liquid T.

At this time, the particles P from which the target substance Q has been removed by the separated liquid T cannot pass through the filter 22, but the target substance Q is discharged together with the separated liquid T to the outside of the filter member 2 and collected. To be done.

The separation liquid T is not particularly limited, but for example, various kinds of water such as distilled water, ion-exchanged water, RO water, pure water and ultrapure water, and various buffers (buffers) adjusted to a predetermined pH. Liquid) and the like.

Incidentally, such a collecting operation may be carried out only once, or may be repeated a plurality of times as required. That is, the recovery operation may be performed at least once. By repeating the recovery operation, the recovery rate of the target product Q can be improved.

When the collecting operation is repeated, the separated liquid T may be replaced without replacement, and the separated liquid T may be replaced with a new separated liquid T for each collecting operation. You may

The object Q can be collected (collected) from the sample S through the above steps.

When one sample processing operation is completed, the tip 1 and the member with filter 2 are moved to the dispensing device 1
It is removed from 0 and discarded. That is, it is preferable that both the chip 1 and the filter-equipped member 2 be disposable after each sample treatment. Accordingly, it is possible to preferably prevent the target object Q obtained from one sample S from being mixed with the target object Q obtained from another sample S (contamination).

The target product Q thus obtained is subjected to post-treatment which is appropriately selected according to its type, purpose and the like. Thereby, for example, qualitative analysis, quantitative analysis, etc. of the target substance Q, concentration, isolation, etc. can be performed.

According to the sample processing method described above, the object Q can be easily and surely collected from the sample S by a simple method using the dispensing device 10 and the chip set. .

Further, since each treatment is carried out in the chip 1 and the member 2 with the filter, a small amount of the sample S, a small amount of the reagent R,
It can be treated with the washing liquid W and the separation liquid T. Therefore, such a sample processing method can be performed at an extremely low cost.

In the sample processing method of this embodiment,
Although the steps [S1] to [S4] are included, the present invention is not limited to this, and the step [S
3] and step [S4], or both of them can be omitted, and a step for any purpose can be added.

Next, another configuration example of the chip 1 will be described with reference to FIG. In the following, the chip 1 shown in FIG. 6 will be described focusing on the differences from the chip 1, and the description of the same items will be omitted.

The chip 1 shown in FIG. 6 has a reduced diameter portion 1a having a reduced outer diameter and inner diameter at approximately the center in the longitudinal direction, and is otherwise the same as the tip 1 described above.

By providing the reduced diameter portion 1a on the chip 1, it becomes possible to process a smaller amount of the sample S.

From this viewpoint, the size (dimension) of the reduced diameter portion 1a is preferably set as follows, for example.

The inner diameter of the reduced diameter portion 1a is preferably about 0.1 to 2 mm, more preferably about 0.3 to 0.7 mm.

In the illustrated construction, the inner diameter and outer diameter of the reduced diameter portion 1a are substantially constant, but they may change continuously or stepwise. In this case, the average inner diameter of the reduced diameter portion 1a may be set to fall within the above range.

The capacity of the reduced diameter portion 1a is 0.1-10.
The amount is preferably about 0 μL, more preferably about 1 to 50 μL, even more preferably about 5 to 10 μL.

In such a chip 1, by appropriately setting the size of the reduced diameter portion 1a, the throughput of the sample can be set to an arbitrary value.

Next, the structure of the processing apparatus 100 provided with the above-mentioned dispensing apparatus 10 will be described with reference to FIG.

FIG. 7 is a schematic diagram showing the overall structure of a processing apparatus equipped with a dispensing apparatus used in the present invention. In the following description, in FIG. 7, the upper side is “upper”, the lower side is “lower”, the left front side of the paper is “front”, and the right back side of the paper is “rear”.
Say

The processing apparatus 100 shown in FIG. 7 comprises a horizontal stage 210 in the front and a vertical stage 220 in the rear.

On the horizontal stage 210, a chip setting part 310 for setting the chip 1, a filter-equipped member setting part 320 for setting the filter-equipped member 2, and various liquids (reagent R, washing liquid W, separation liquid T) are stored. Container setting section 330 for setting the storage container, a tube for storing the sample, and a processing tube used for various kinds of processing (for example, the above-mentioned container X
Etc.) and the tube installation part 340.

Further, on the vertical stage 220, the dispensing device 10 is provided with an x-axis direction moving mechanism 410 capable of moving in the x-axis direction (horizontal direction in FIG. 7) and a y-axis direction (oblique front-back direction in FIG. 7). ), And a z-axis direction moving mechanism 4 capable of moving in the z-axis direction (vertical direction in FIG. 7).
It is installed via a moving mechanism 400 composed of 30 and 30.

Further, the processing device 100 is electrically connected with a control means such as a personal computer (not shown), and the operation of each part of the processing device 100 is controlled by the control means. .

In the processing apparatus 100 as described above, the dispensing mechanism 10 is moved in the three-dimensional direction by the operation of the moving mechanism 400, and the tip 1 is attached to the nozzle 30, the member 2 with a filter is attached to the tip 1, Suction and discharge of various liquids (processing in the steps [S1] to [S4]) are performed.

As described above, in the step [S1], when the mixed solution of the sample S and the reagent R is heated, the lower part of the tube setting section 340 where the container X used for carrying operation is set. At one or both of the predetermined positions of the dispensing device 10, for example, a heating unit (temperature adjusting unit) configured by a Peltier element or the like may be provided.

Further, as described above, in the step [S1], when the mixed liquid of the sample S and the reagent R is irradiated with electromagnetic waves, for example, the container X of the tube setting section 340 used for carrying operation is installed. An electromagnetic wave irradiating means capable of generating and irradiating an electromagnetic wave (microwave, ultraviolet ray) may be provided below the portion.

In the above description, the case where the sample is processed by using the pipetting device (automating device) has been described. However, in the present invention, the sample is processed manually by using, for example, a pipette (micropipetter). It can also be applied when

The sample processing method and chip set of the present invention have been described above based on the illustrated embodiments, but the present invention is not limited to these.

For example, in the chip set of the present invention, the configuration of each part can be replaced with any configuration capable of exhibiting the same function.

When a dispensing device having a plurality of nozzles is used, a plurality of specimens can be processed at the same time.

Further, for example, a plurality of types of filter-equipped members each having a filter having at least one of physical properties and chemical properties different from each other are prepared, and these members are sequentially replaced or used at the same time as the filter-attached member. By doing so, it is possible to collect (collect) various kinds of target substances by one-time sample processing.

[0114]

As described above, according to the present invention, it is possible to easily and surely collect an object from a sample without requiring a large-scale device.

Since each treatment is carried out in the chip or the member with the filter, a small amount of sample can be treated. In addition, the sample processing method of the present invention can be implemented at low cost.

[Brief description of drawings]

FIG. 1 is a schematic view showing a step (step [S1]) of an embodiment of a method for treating a sample of the present invention.

FIG. 2 is a schematic view showing a step (step [S2]) of the embodiment of the sample processing method of the present invention.

FIG. 3 is a schematic view showing a step (step [S3]) of the embodiment of the sample processing method of the present invention.

FIG. 4 is a schematic view showing a step (step [S4]) of the embodiment of the sample processing method of the present invention.

FIG. 5 is a partial cross-sectional view showing a configuration of a chipset of the present invention and a dispensing device used in the present invention.

FIG. 6 is a cross-sectional view showing another configuration example of the chip.

FIG. 7 is a schematic diagram showing an overall configuration of a processing apparatus including a dispensing device used in the present invention.

[Explanation of symbols]

1 chip 1a reduced diameter part 2 Filter-equipped member 21 Tubular member 22 filters 221 pores 10 dispensing device 20 dispensing pump 30 nozzles 31 passage S sample Q target O unnecessary material R reagent P particles W cleaning liquid T separation liquid X container 100 processing equipment 210 horizontal stage 220 vertical stage 310 Chip installation section 320 Filter installation part 330 Container installation section 340 tube installation section 400 moving mechanism 410 x-axis direction moving mechanism 420 y-axis direction moving mechanism 430 z-axis movement mechanism

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) // G01N 33/48 G01N 1/28 YF term (reference) 2G045 BA01 HA01 2G052 AA29 AA30 AA32 AA33 AB18 AB19 AB20 AD29 AD46 CA03 CA20 CA28 CA33 EA03 EB11 ED04 ED05 ED11 FB02 FB09 FC05 FC11 FC15 JA04 JA16 2G058 EA02 ED35 FA04

Claims (22)

[Claims] 1. A tip mounted on the tip of a nozzle,
Aspirate a sample and a reagent containing particles capable of supporting an object, and then, a tubular member that can be attached to the tip of the chip,
A filter-equipped member that is provided in the flow path of the tubular member and that includes a filter that can capture the particles is attached to the tip of the chip, and a mixed liquid of the sample and the reagent is supplied from the inside of the chip. A method for treating a specimen, wherein the particles carrying the target substance are captured by the filter by discharging. 2. The method for treating a sample according to claim 1, wherein after the sample and the reagent are sucked into the chip, an operation of stirring the mixed liquid of the sample and the reagent is performed. 3. The sample processing method according to claim 2, wherein the stirring operation is performed by discharging the mixed liquid from the inside of the chip and then again sucking the mixed liquid into the chip at least once. . 4. The method for treating a sample according to claim 1, wherein the particles are allowed to carry the target substance while heating a mixed liquid of the sample and the reagent. 5. The method for treating a sample according to claim 4, wherein the heating temperature is 20 to 100 ° C. 6. The method for treating a specimen according to claim 1, wherein the particles are made to carry the target substance while irradiating a mixed liquid of the specimen and the reagent with electromagnetic waves. 7. The method for treating a sample according to claim 6, wherein the electromagnetic wave is a microwave. 8. The method for treating a sample according to claim 6, wherein the electromagnetic wave is ultraviolet light. 9. The method for treating a specimen according to claim 1, wherein at least the surface of the particle is solid-phased with a substance capable of supporting the target substance. 10. The average particle size of the particles is 10 to 100.
The method for treating a specimen according to any one of claims 1 to 9, which has a thickness of 0 μm. 11. The object is obtained by capturing the particles carrying the target substance with the filter, sucking the cleaning liquid into the member with the filter, and then discharging the cleaning liquid at least once. The method for treating a specimen according to any one of claims 1 to 10, wherein the particles carrying a substance are washed. 12. After the particles carrying the target substance are captured by the filter, the particles are placed in the member with a filter.
A method in which the target liquid separated from the particles is recovered in the separation liquid by suctioning a separation liquid that separates the target product from the particles, and then performing an operation of discharging the separation liquid at least once. The method for treating a sample according to any one of 1 to 11. 13. The sample processing method according to claim 1, wherein the filter has a property of not substantially adsorbing the target substance. 14. The method for treating a specimen according to claim 1, wherein the tubular member has a shape substantially equal to the shape of the tip. 15. The method for treating a specimen according to claim 1, wherein the tip has a reduced diameter portion in which an outer diameter and an inner diameter are reduced in a substantially central portion in a longitudinal direction thereof. 16. The method for treating a sample according to claim 15, wherein the inner diameter and the outer diameter of the reduced diameter portion are substantially constant. 17. The inner diameter (average) of the reduced diameter portion is 0.1.
The method for treating a specimen according to claim 15 or 16, wherein the specimen has a size of ˜2 mm. 18. The volume of the reduced diameter portion is 0.1 to 100.
18. The method for treating a sample according to claim 15, which is μL. 19. The sample processing method according to claim 1, wherein both the chip and the filter-equipped member are disposable after each sample processing. 20. The method for treating a specimen according to claim 1, wherein the target substance is a microorganism, nucleic acid, protein, hormone, amino acid, peptide or nucleotide. 21. The processing of the sample is performed by suctioning and transferring the sample into the chip by operating a dispensing device equipped with a dispensing pump for sucking and dispensing a liquid. The method for processing a sample according to Crab. 22. A chipset used in the method for treating a sample according to claim 1, wherein the chip is attachable to a tip of a nozzle of a dispensing device, and is attached to a tip of the tip. A chipset having a filterable member including a possible tubular member and a filter provided in a flow path of the tubular member.