WO2017065585A1 - Method for selectively treating biological sample - Google Patents

Abstract

A method for selectively treating a biological sample is provided, the method comprising the steps of: providing a substrate on which biological samples are arranged; decoding types and locations of the biological samples and then selecting at least one sample to be tested among the biological samples; forming a barrier structure to surround the periphery of the sample to be tested and thus partitioning an area in which the sample to be tested is located, thereby spatially separating the sample to be tested from the remaining biological samples; introducing a biochemical reaction reagent within the partitioned area; and performing a biochemical reaction between the biochemical reaction reagent and the sample to be tested.

Description

Selective treatment method of biological sample

The technology disclosed herein relates to a method for selectively treating a biological sample, and more particularly, to select a desired target sample from a biological sample and to selectively process and recover a biological sample without contamination of other biological samples. It is about.

As an example of a biological sample, tissue consists of a very large number of cells and not all of them have the same DNA or RNA. Therefore, in order to accurately determine which tissues, for example, cancerous tissues have abnormalities, a technique for separating cells one by one and analyzing them as single cells is required.

Single cell analysis is divided into cell observation method and DNA / RNA / protein analysis method, where the cell observation method is different from DNA / RNA / protein analysis method that separates analyte from the beginning for analysis. During the observation, the target cells selected for analysis are separated later, in which case there is a risk of damage or contamination of the target cells. In addition, there is a disadvantage that it is difficult to separate several cells at once and difficult to analyze high purity at a small amount.

Commercially available techniques of single cell analysis include a method of separating cells one by one in a suspended cell state, and a method of separating cells using microdissection on solid tissues.

In the former case, a microfluidics device is disclosed in US Patent Publication No. 2013-0295602, and a technology related to Fluorescence-activated cell sorting (FACS) is disclosed in US Patent Publication No. 2008-0124726. In addition, micropipetting may be included, but only cells floating in a liquid state can be analyzed one by one.

In the latter case, there is a laser microdissection, which mainly involves the separation of solid-state cells with a laser, which can lead to damage to the cells, less accuracy in separating only one cell, and is difficult to automate and multiple at once. There is a disadvantage in that it is difficult to separate the cells. In particular, there is a problem that the target sample is contaminated because it corresponds to a method of separating the target sample and moving to the individual test tube.

According to one aspect of the technology disclosed herein, providing a substrate on which biological samples are disposed; Selecting at least one target sample of the biological samples by reading the type and location of the biological samples; Spatially separating the target sample from the remaining biological samples by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing a biochemical reagent into said compartmentalized region; And performing a biochemical reaction between the biochemical reaction reagent and the target sample.

According to another aspect of the technology disclosed herein, there is provided a method comprising providing a substrate on which biological samples are disposed; Observing the biological samples and selecting at least one target sample of the biological samples; Spatially separating the target sample from the remaining biological samples by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing the biochemical reaction reagent into the compartmentalized region; Pretreating the target sample through a biochemical reaction between the biochemical reaction reagent and the target sample; And analyzing the target sample on the substrate or recovering and analyzing the sample from the substrate.

According to another aspect of the technology disclosed herein, there is provided a method comprising providing a substrate on which biological samples are disposed; Selecting at least one target sample of the biological samples by reading the type and location of the biological samples; Forming a microwell in which the target sample is located by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing a lysis solution into the microwell to dissolve the sample of interest; Preparing a library of nucleic acid molecules for sequencing by pretreating nucleic acid molecules derived from the target sample by the lysis with a biochemical reagent; Recovering the library of nucleic acid molecules from the substrate; And sequencing the recovered library of nucleic acid molecules.

According to another aspect of the technology disclosed herein, a stage comprising a substrate on which biological samples are placed; An imaging processor for observing the biological samples and selecting at least one target sample; A microwell manufacturing unit for forming a microwell by stacking a barrier structure to surround a region around the selected target sample; And a biochemical treatment unit for biochemical reaction of the target sample in the microwell.

1 is a process flow diagram showing an embodiment of a method for selectively processing a target sample in a biological sample.

2 is a schematic diagram illustrating an embodiment of a process for forming a barrier structure for screening and analyzing a target sample in a biological sample.

3 is a partially enlarged cross-sectional view of a substrate on which a barrier structure corresponding to C of FIG. 2 exists.

 4 illustrates an embodiment of introducing a biochemical reaction reagent onto a substrate on which a barrier structure is formed.

FIG. 5 is an example of a specific application of the method for selectively treating a biological sample shown in FIG. 1.

6 is a view showing an embodiment of a selective treatment apparatus for a biological sample.

Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. The embodiments described below are provided as examples to sufficiently convey the spirit disclosed to those skilled in the art. Thus, the disclosed technology is not limited to the embodiments described below and may be embodied in other forms. And in the drawings, the width, length, thickness, etc. of the components may be exaggerated for convenience. Like reference numerals denote like elements throughout the specification. The overall description has been made at the point of view of an observer, and when one component is on another component, this includes not only the case where the component is directly above the other component but also another component in the middle.

1 is a process flow diagram showing an embodiment of a method for selectively processing a target sample in a biological sample. Referring to FIG. 1, in step 110, a substrate on which biological samples are disposed is provided.

The biological sample may or may not be fixed on the substrate. The biological sample may be at least one selected from the group consisting of tissues, blood, cells, DNA, RNA, proteins, exosomes, metabolites, and biopsy specimens, unless otherwise specified. The tissue may be, for example, solid tissue, and the cell may be, for example, suspension cells or smeared cells.

The substrate is not particularly limited so long as it provides a surface for supporting the biological sample, but in a crowd consisting of slide glass, microbeads, nanoparticles, nanostructures, capillaries, microfluidic supports, pore structures, sponge structures, and dendrimers It may be one or more selected. The substrate may typically be slide glass.

The substrate may be made of glass, silicon or a polymer material.

In one embodiment, the substrate may be a functionalized substrate modified with one or more selected from the group consisting of DNA, RNA, proteins, antibodies, chemicals. For example, it may be a microarray substrate or a supercapacity sequencing substrate in which biological samples such as DNA or protein are integrated.

In operation 120, at least one target sample of the biological samples is selected by reading the type and location of the biological samples.

The location of the biological samples can be read, for example, through image observation, fluorescence signals or coordinate information. The biological sample is not limited as long as it is a staining technique that can obtain information of the biological sample. It can be dyed by one dyeing technique to provide image information. In some embodiments, the step of screening the subject sample may be performed by obtaining positional information of the biological samples in a manner that is directly observed through an optical microscope or an electron microscope or in an automated manner using separate software. For example, after the image of the biological sample is obtained, the sample may be divided into several or dozens of groups using clustering or classification techniques in an automated manner, and then the sample may be automatically or manually selected.

In operation 130, a barrier structure is formed to enclose a periphery of the target sample to partition the region in which the target sample is located to spatially separate the target sample from the remaining biological samples.

2 is a schematic diagram illustrating an embodiment of a process for forming a barrier structure for screening and analyzing a target sample in a biological sample. Referring to FIG. 2, side cross-sectional views of cut lines indicated by dotted lines in the left plan view of each of A, B, C, and D pictures are written together in the right view. Specifically, FIG. 2A schematically illustrates an image of a biological sample 220 disposed on a substrate 210, for example, a plurality of cells, and FIG. 2B illustrates an example sample 220 selected on a substrate. Single cells are shown. In FIG. 2C, a barrier structure 230 surrounding the periphery of the target sample 220 is formed. The barrier structure 230 is introduced to overcome the problem of damaging the target sample in any way while separating the target specimen from the substrate and not damaging the target sample. Referring to FIG. 2D, the barrier structure 230, for example, prevents the biochemical reaction reagent 270 from flowing out of the partitioned region 260 in which the target sample 220 is located on the substrate 210. Block the effect of reagent 270 on other samples. The barrier structure 230 may be made of a material that is physically or chemically protected with respect to the biochemical reaction reagent 270. For example, the barrier structure 230 may be made of at least one material selected from the group consisting of polymer resin, wax, metal, metal oxide, and glass.

In one embodiment, forming the barrier structure may include applying a curable material entirely on the substrate, curing with light or heat, and then removing the uncured residual curable material. In another embodiment, forming the barrier structure may include dispensing the curable material along the shape of the barrier structure and curing with light or heat. In another embodiment, forming the barrier structure may include providing a curable material through a microfludic chip or through a microfluidic chamber. When the microfluidic technology is used to provide a curable material, the height of the barrier structure may be limited, the curable material may be uniformly provided for the entire substrate, and the consumption of the curable material may be reduced.

The curable material may include unsaturated monomers, non-limiting examples of unsaturated monomers include ethoxylated trimethylolpropane triacrylate, curable epoxy (product name NOA), polyethylene glycol diacrylate, polypropylene glycol diacrylate, Polyurethane acrylates and combinations thereof. For example, the above-described polyethylene diacrylate has a structure having acrylate functional groups at both ends of polyethylene glycol, so that free radical polymerization can be crosslinked into a three-dimensional structure. In addition, the curable material may include any form of medium that can change from a liquid to a solid.

The curable material may further include an initiator and may cause free radical polymerization by an external energy source. The initiator may be an azo compound or a peroxide. The curable material may further include a suitable crosslinking agent, and examples thereof include N, N'-methylenebisacrylamide, methylenebismethacrylamide and ethylene glycol dimethacrylate. Suitable energy sources for the curing reaction may include, without limitation, heat, ultraviolet light, visible light, infrared rays and electron beams.

The curable material may be a material that bonds to the substrate simultaneously with curing. For example, during hardening, the material forms a bond with the glass, which allows the barrier structure to be more strongly secured to the substrate.

The curable material may penetrate into a biological sample and be cured. For example, a basic dissolving reagent or proteinase may be mixed with a general curable material so that the curable material may penetrate between or inside tissues and then be cured by an external energy source.

In order to increase the permeability of the curable material to the biological sample, a process of pretreating the biological sample may be included before supplying the curable material. For example, a chemical reaction that creates pores in the cell membrane can increase the permeability of the curable material. As another example, the curable material may be provided after leaving only the cell nucleus in a biological sample through a biochemical reaction that dissolves the cell membrane but does not dissolve the nuclear membrane.

The method for forming the barrier structure may be performed by various patterning processes, and for example, may be formed by a lithography method, an inkjet printing method, or a 3D printing method. Specifically, in order to form the barrier structure, general optical lithography may be used, or maskless ligthography and laser scanning using a digital mirror device (DMD) may be used.

The lithography method for forming the barrier structure may be a method of performing a lithography in a large area without a lens between a mask and a substrate, and may be a method of sequentially performing lithography on various regions at high resolution using a lens. In particular, in the case of a lithography using a lens, even if the biological sample is changed and the target region is changed, the barrier structure is sequentially formed using one or several kinds of fixed masks, thereby eliminating the need to change the mask every time the sample is changed.

The method of sequentially performing a lithography on a biological sample using the lens may further include an optimization algorithm for forming a barrier structure on a desired sample region using one or several kinds of fixed masks. The method may further include an optimization algorithm for forming a barrier structure on a desired target sample region using a digital mirror device (DMD).

Lithography using the one or several kinds of fixed masks may further include a method of controlling the size and resolution of the barrier structure by adjusting the magnification of the lens.

As an example of optical lithography, the method for forming the barrier structure may be a method of forming a barrier structure structured by an optical lithography method using a mask in which a patterned pattern is engraved. For example, the standardized pattern may be a lattice masking pattern. This method does not form the barrier structure only around the selected target sample, but the standardized barrier structure may be formed to include the target sample, and the barrier structure forming process is simpler than the maskless lithography.

The barrier structure may be formed by supplying a hydrophilic coating or a hydrophobic coating. In a preferred embodiment, a hydrophobic coating agent may be provided around the target sample to form a barrier structure such that an aqueous solution may be supplied only to the inside of the barrier structure.

The area partitioned by the barrier structure may be outside or inside the closed curve created by the barrier structure. For example, by forming a barrier structure covering the biochemical sample in a circular shape, the target sample may be positioned outside the circle, and the target sample may be positioned inside the circle.

The barrier structure may have a microwell shape. In this case, the region partitioned by the barrier structure may be inside or outside the microwell. The microwells may have a width within a well of 1 μm to 1 mm, and the shape of the wells may be circular, polygonal, or amorphous.

The method for forming the barrier structure may be a method of placing or assembling an already formed structure on a substrate. For example, a barrier structure may be formed by assembling a microwell structure or micro particles on a substrate.

One or more partitioned regions may exist on the substrate, and in the case of a plurality of partitioned regions, their sizes may be the same or different. The size of the compartmentalized region may be determined according to the number of target samples of interest, and one or more target samples may be present in the compartmentalized region, for example, types of target samples and Depending on the size may be dozens or hundreds or more in the partitioned area. Also, for example, if the subject sample is a cell, the compartmentalized region may be sized to receive one cell or even smaller than one cell. That is, the compartmentalized region may be present on a portion of one cell.

3 is a partially enlarged cross-sectional view of a substrate on which a barrier structure corresponding to C of FIG. 2 exists. Referring to FIG. 3, when a specific single cell 310 is selected as a target sample, the barrier structure 320 is formed surrounding the specific single cell 310, so that only one cell in the partitioned region 330 is formed. The other cells present and external suggest a form that is spatially separated from the subject sample. Thus, in this state, the biological sample can be later analyzed at the single cell level.

Since the formation of the barrier structure does not significantly affect the biological sample, the biological sample may be cultured after the barrier structure is formed. For example, by forming a barrier structure selectively around cells having a desired phenotype and then culturing the cells, the cells can be cultured by separating the cells with other phenotypes.

In step 140, a biochemical reagent is introduced into the compartmentalized region.

Referring to FIG. 2D, the biochemical reagent 270 is introduced into the partitioned region 260 in which the selected target sample 225 is located. The biochemical reaction reagent may be introduced using a microdispensing method. The microdispensing is a technique for introducing a liquid medium of less than 1 microliter, or less than 0.1 microliter. Preferred examples of the microdispensing include inkjet printing. If necessary, the introduction of the biochemical reagent may be performed by pipetting of 0.1 microliter or more.

The biochemical reagent may also be supplied after assembling the microfluidic chip or microfluidic chamber onto a substrate. Feeding biochemical reagents using microfluidic technology can reduce reagent consumption and prevent reagent evaporation. If you want to provide two or more types of biochemical reagents sequentially, it can be easily provided by reassembling a larger microfluidic chamber.

The biochemical reagents include a lysis solution, a reagent for PCR reaction, a reagent for whole genome amplification and a reagent for whole transcriptome amplification, reverse transcription, and reverse transcription PCR. (RT PCR), in vitro transcription, rolling circle amplifciation, bisulfite treatment, DNA extraction, RNA extraction, protein extraction, genome editing, accessible Reagents required for various biochemical reactions, such as permeabilization, cell in situ sequencing, can be used alone or in combination.

As the lysis solution, for example, a basic dissolution reagent or proteinase (Proteinase) may be used.

At this time, before introducing other biochemical reaction reagents, use a lysis solution and then introduce other biochemical reaction reagents according to the purpose or introduce the lysis solution and other biochemical reaction reagents together into the compartment. You may.

The biochemical reagent is a library preparation reaction for massively parallel sequencing including, for example, transposase, ligase, fragmentation, etc. It may be a reagent for.

The biochemical reaction reagent may be injected only within the area partitioned by the barrier structure, and if necessary, may be injected in addition to the area partitioned by the barrier structure as well as the partitioned area.

4 illustrates an embodiment of introducing a biochemical reaction reagent onto a substrate on which a barrier structure is formed. Referring to FIG. 4, even if the biochemical reaction reagents are injected not only in the compartmentalized region but also in the region outside the compartmentalized region, if the reaction results are not mixed with each other due to the effect of preventing diffusion by the barrier structure, the target sample may be precisely determined. Analysis may be possible. For example, the DNA of a cell is a very heavy polymer that tends to spread and remain in place rather than beyond the barrier structure, even when biochemical reactions proceed.

Before introducing the biochemical reagent, the biochemical reagent may be introduced after covering the hydrogel on the biological sample in which the barrier structure is formed in order to minimize the diffusion of the biochemical material. For example, agarose may be introduced onto a biological sample on which a barrier structure is formed, and then hardened to form a hydrogel, and biochemical reaction reagents may be introduced thereon. As another example, the biochemical reagent may be introduced by soaking the biochemical reagent in the hydrogel and then covering the hydrogel on the biological sample.

The biochemical reaction reagent may be used to inject a reagent for reacting a biomaterial on a DNA microarray substrate, a high capacity sequencing plate, or a flow cell for ultra high capacity sequencing.

The biochemical reagent may be a decrosslink reagent that releases the crosslink of the curable curable material to form a barrier structure. For example, when the barrier structure covers the area where the sample is located, all the biological samples around the area where the barrier structure is present are dissolved and removed, and then the decrosslink reagent is supplied to expose the sample and further biochemical reaction is performed. Can carry out

In step 150, the target sample is pretreated through the biochemical reaction of the biochemical reaction reagent and the target sample. These biochemical reactions result in amplification of nucleic acid molecules derived from a target sample, for example.

The biochemical reaction may include a reaction in which different kinds of oligonucleotides are ligated and inserted into an existing reactant to tag the reactants of different microwells. . The tagging method also includes beads, microparticles, hydrogels, droplets, or core-shell particles having different types of oligonucleotides attached thereto. It may include a method of injecting. The tagging method may also include a method of assembling a microarray substrate to which oligonucleotides are attached and a substrate on which biological samples are placed. The biochemical reaction may also include pretreatment reactions for ELISA, aptamer binding, mass spectroscopy.

In one embodiment, the biochemical reaction may be further promoted by applying a physical force such as stirring, vibration, or ultrasonic waves to the partitioned region. In addition, depending on the purpose, it may be necessary to control the temperature to induce an appropriate biochemical reaction. On the other hand, it is preferable to add a drying prevention treatment process, such as anti-humidity maintenance and sealing with oil in the pre-treatment process can prevent the drying of the biochemical reaction reagents.

In step 160, the target sample is recovered from the substrate.

The process of recovering the target sample from the substrate may be performed by one or more methods selected from the group consisting of micro pipetting, ultrasound, light pressure, laser, stamping, pulling, and absorption and hybridization using microbeads. As a specific example of the recovery method, one or more target samples may be grouped, pooled and recovered after indexing, and the target samples in each compartment may be recovered using a microfluidic pipe.

As another preferable example of a recovery method, a biochemical reaction may be performed on a sample of a desired type by forming a barrier structure only on a sample of a desired type in a biological sample having various types of samples, and then pooling the sample. . For example, if a barrier structure is formed only around cancer cells in a tissue mixed with normal cells and cancer cells, only the information about the cancer cells can be obtained by pooling without the need for barcode tagging.

Selective treatment method of a biological sample disclosed herein can be screened while observing a desired target sample from the biological sample through imaging and the like, and can be pretreated without being contaminated with other biological samples, and easily removes the desired target sample from the substrate without damage. It can be recovered.

According to another aspect of the technology disclosed herein, a method for selectively analyzing a biological sample on a substrate by analyzing the target sample obtained by the above-described method of steps 110 to 150 on the substrate as it is or recovered from the substrate Is provided.

One embodiment of a method for selectively analyzing a biological sample may be performed as follows. First, a substrate on which biological samples are disposed is provided. Next, the biological samples are observed to select at least one target sample from the biological samples. Subsequently, a barrier structure is formed to surround the target sample to partition the region in which the target sample is located to form a microwell in which the target sample is located. And a lysis solution is introduced into the microwell to dissolve the target sample. Next, the nucleic acid molecules derived from the target sample by the lysis are pretreated with a biochemical reagent to prepare a library of the nucleic acid molecules for sequencing. The library of nucleic acid molecules is then recovered from the substrate. The biological samples on the substrate can then be selectively analyzed by sequencing the recovered library of nucleic acid molecules.

Preferably, the sequencing may be performed by a massively parallel sequencing technique such as Next Generation Sequencing, which is very efficient.

By performing the sequencing, optical and electromagnetic signals sequentially generated according to the type of the nucleotide sequence may be provided together with the location information. The ultra-high-capacity sequencing method analyzes hundreds of thousands of sequences at the same time, so that the analysis throughput is very high-throughput compared to the conventional sequencing method (statistics for the sequencing of the sample to be analyzed) Resources may be provided.

FIG. 5 is an example of a specific application of the method for selectively treating a biological sample shown in FIG. 1. Referring to FIG. 5, the method for selectively treating a biological sample according to the technology disclosed herein may be performed in the following order.

Referring to FIG. 5A, at least one target sample of the biological samples is selected by observing a substrate on which cells, which are a kind of biological sample, are disposed. A UV curable polymer is applied thereon and irradiated with a UV lamp through a digital mirror device (DMD) around a selected sample through an imaging device (not shown). Photocuring the curable material using. Referring to FIG. 5B, a barrier structure is formed to surround the object sample so as to exist in the microwell in which the object sample is located. Referring to FIG. 5C, a library of nucleic acid molecules derived from the target sample is obtained by introducing a biochemical reaction reagent such as a lysis solution into the microwell using an inkjet dispenser and pretreating the target sample. Referring to FIG. 5 (d), the obtained library of nucleic acid molecules is recovered and sequenced using a massively capacity sequencing machine.

According to the selective treatment method or analysis method of the biological sample as described above, it is possible to determine exactly which side or abnormality the tissue (especially cancer tissue) has.

For example, cancer tissues extracted from cancer patients may be coated on glass, and then stained with a commonly used staining reagent (eg, giemsa), and the desired cells may be selected and collected while observing under a microscope, for sequencing.

 Compared with the laser microcutting technique, since the laser is not used, the problem of contamination is lessened because it does not damage the biological sample and does not need to move the biological sample to another reaction space for the biochemical reaction. In addition, due to the nature of the process, automation is easy, so commercialization is easy.

In particular, since a biochemical reaction on a biological sample occurs in a micro-scale reaction space, only a small amount of reagent may be used, and there is an advantage in that mass analysis is easy.

According to another aspect of the technology disclosed herein, an apparatus for use in a method for selectively treating a biological sample is provided. 6 is a view showing an embodiment of a selective treatment apparatus for a biological sample. Referring to FIG. 6, the apparatus 600 for selectively processing a target sample among biological samples includes a stage 610 on which a substrate on which biological samples are disposed is mounted; An imaging processor 620 for observing the biological samples and selecting at least one target sample; A microwell manufacturing unit 630 for forming a barrier structure to surround a region around the selected target sample; And a biochemical treatment unit 640 for biochemical reaction of the target sample in the microwell.

The stage 610 may include a mount for mounting a substrate, and is preferably an electric stage for precise manipulation.

The imaging processor 620 may include an imaging device for reading the type and position of a target sample on a substrate, and specific examples thereof include an optical microscope or an electron microscope for observation.

The microwell manufacturing unit 630 may include, for example, a lithography system, an inkjet printing system, or a 3D printing system. For example, the microwell manufacturing unit 630 may include a photofluidic maskless lithography system. To this end, the microwell manufacturing unit 630 may include a UV light source, a digital mirror device, a lens, and the like.

The biochemical treatment unit 540 may include a storage means and a supply means of a biochemical reaction reagent. In one embodiment, the biochemical processor 540 may include a reaction promoting device for applying a physical force such as stirring, vibration, or ultrasonic waves to the partitioned reaction space in which the target sample is located, such as a microwell. Alternatively, in one embodiment, the biochemical processor 540 may include a temperature control device for controlling the reaction temperature.

The above-described selective processing apparatus of the biological sample may further include an extraction apparatus for extracting a target sample pretreated with a biochemical reaction reagent and / or a sequencing apparatus for sequencing the sample extracted by the extraction apparatus, if necessary. have. The sequencing equipment is not specified as long as it is a sequencing equipment that is commonly used, but it may be desirable to use a massively parallel sequencing sequencing in consideration of the analysis efficiency of sequencing.

Selective processing apparatus for a biological sample according to an embodiment of the present disclosure can be applied to a variety of applications because it can be analyzed up to a single cell level. That is, the present invention may be applied to a bio analyzer or a gene diagnostic equipment for selectively analyzing a desired biological sample.

Although the embodiments of the technology disclosed in detail above have been described in detail, those of ordinary skill in the art may modify the technology disclosed herein without departing from the spirit and scope of the technology disclosed herein. It will be understood that it can be carried out by.

Claims (17)

Providing a substrate on which biological samples are disposed; Selecting at least one target sample of the biological samples by reading the type and location of the biological samples; Spatially separating the target sample from the remaining biological samples by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing a biochemical reagent into said compartmentalized region; And Selective treatment method of a biological sample comprising the step of performing a biochemical reaction of the biochemical reaction reagent and the target sample. The method of claim 1, The biological sample is selected from the group consisting of tissue, blood, cells, DNA, RNA, protein, exosomes, metabolites (metabolite) and biopsy specimen (bipsy specimen) selective treatment method of a biological sample. The method of claim 1, Selecting the target sample is a method of selectively processing a biological sample is performed by obtaining the position information of the biological sample in a direct observation via an optical microscope or an electron microscope or in an automated manner using a separate software. The method of claim 1, The step of reading the position of the biological sample is a selective analysis method of the biological sample is performed through image observation, fluorescence signal or coordinate information. The method of claim 1, Wherein said partitioned region has a microwell shape. The method of claim 1, And the barrier structure is made of a material that is physically or chemically protected so that the biochemical reaction reagent does not flow out of the compartmentalized region where the target sample is located. The method of claim 6, The barrier structure is a selective treatment method of a biological sample consisting of at least one material selected from the group consisting of polymer resin, wax, metal, metal oxide and glass. The method of claim 1, The barrier structure is formed by a method selected from the group consisting of a lithography method, inkjet printing, and a 3D printing method. The method of claim 1, Forming the barrier structure comprises applying a curable material onto the substrate and curing with light or heat. The method of claim 1, And recovering the target sample from the substrate. The method of claim 1, Introduction of the biochemical reagent is a selective treatment method of a biological sample is carried out using a microdispensing method. The method of claim 1, The biochemical reaction reagent is at least one selected from the group consisting of a lysis solution, a reagent for PCR reaction, a reagent for whole genome amplification, and a reagent for whole transcriptome amplification. Selective treatment of biological samples. The method of claim 10, The step of recovering the target sample from the substrate is a selective processing method of a biological sample is performed by at least one method selected from the group consisting of micro pipetting, ultrasonic waves, light pressure, laser, stamping, pooling and absorption. Providing a substrate on which biological samples are disposed; Observing the biological samples and selecting at least one target sample of the biological samples; Spatially separating the target sample from the remaining biological samples by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing the biochemical reaction reagent into the compartmentalized region; Pretreating the target sample through a biochemical reaction between the biochemical reaction reagent and the target sample; And Selective analysis method of the biological sample comprising the step of analyzing the target sample on the substrate or recovered from the substrate. Providing a substrate on which biological samples are disposed; Observing the biological samples and selecting at least one target sample of the biological samples; Forming a microwell in which the target sample is located by forming a barrier structure to surround the target sample and partitioning an area in which the target sample is located; Introducing a lysis solution into the microwell to dissolve the sample of interest; Preparing a library of nucleic acid molecules for sequencing by pretreating nucleic acid molecules derived from the target sample by the lysis with a biochemical reagent; Recovering the library of nucleic acid molecules from the substrate; And And sequencing the recovered library of nucleic acid molecules. A stage having a substrate on which biological samples are disposed; An imaging processor for observing the biological samples and selecting at least one target sample; A microwell manufacturing unit for forming a microwell by stacking a barrier structure to surround a region around the selected target sample; And And a biochemical treatment unit for biochemical reaction of the target sample in the microwell. The method of claim 16, The biochemical treatment unit further comprises a reaction promoting device for applying any one of the physical force selected from the group consisting of agitation, vibration and ultrasound.

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