Sample recovery method for microdissection and solid transfer body

Abstract

The embodiment of the invention provides a sample recovery method for microdissection and a solid transfer body. The sample recovery method for microdissection comprises the following steps that a solid transfer body of a sample is manufactured, and the solid transfer body at least comprises a PDMS solid matrix and a solid transfer film; carrying the sample through the solid transfer film, so that the sample is attached to the solid transfer body; and under the condition that the sample needs to be transferred, the attachment of the sample and the solid transfer body is removed, so that the sample meeting the requirement completely falls off to a target area from the solid transfer body. According to the sample recovery method provided by the technical scheme, the solid transshipment film is additionally arranged on the PDMS solid substrate, and the solid transshipment film carries the sample, so that the adsorption recovery rate of the PDMS solid substrate to the cut sample is remarkably improved, and the problem that the sample is difficult to transfer from the PDMS solid substrate to the collection pipe is solved; the requirements on efficient and accurate sample recovery in the field of biological sample research and analysis are met.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological sample collection and processing, and in particular relates to a sample recovery method and a solid transfer carrier for microdissection. Background Art

[0002] In the field of biological sample collection and processing, microdissection technology is often used to obtain specific samples from complex tissues or cell populations. However, existing technologies present numerous challenges in the sample recovery process, such as low sample recovery rates and poor integrity of recovered samples, which severely impact subsequent experimental analysis. These issues limit the further application of microdissection technology in biomedical research, clinical diagnostics, and other fields, and require urgent resolution. Summary of the Invention

[0003] In view of the above problems in the prior art, an object of the embodiments of the present invention is to provide a sample recovery method and a solid transfer carrier for microdissection.

[0004] The technical solution adopted in the embodiment of the present invention is a method for recovering a sample from microdissection, comprising the following steps:

[0005] Making a solid transfer carrier for the sample, wherein the solid transfer carrier at least comprises a PDMS solid matrix and a solid transfer membrane;

[0006] Carrying the sample through the solid transfer membrane so that the sample is attached to the solid transfer carrier;

[0007] When the sample needs to be transferred, the sample is released from the solid transfer carrier so that the sample that meets the needs falls off completely from the solid transfer carrier to the target area.

[0008] Furthermore, the solid transfer carrier for making the sample, which at least includes a PDMS solid matrix and a solid transfer film, includes:

[0009] A film-forming solution is prepared; the film-forming solution is evenly spread on the PDMS solid substrate; and the film-forming solution is dried to form a solid transfer film on the PDMS solid substrate.

[0010] Furthermore, the PDMS solid matrix is ​​pretreated before preparing the solid transfer carrier of the sample, including:

[0011] The PDMS solid substrate is subjected to a hydrophilic treatment to enhance the hydrophilicity of the surface of the PDMS solid substrate and improve the bonding strength between the solid transfer membrane and the PDMS solid substrate;

[0012] The hydrophilic treatment includes at least acid treatment;

[0013] The acid treatment comprises:

[0014] The hydrogen ions in the acid react with the silicon-oxygen bonds on the surface of the PDMS solid substrate, partially breaking them and introducing hydroxyl groups to form hydrophilic silanol groups;

[0015] At the same time, the surface of the PDMS solid substrate is etched with acid to increase the surface roughness of the PDMS solid substrate.

[0016] Furthermore, the hydrophilic treatment further comprises:

[0017] After the PDMS solid substrate is acid-treated, the PDMS solid substrate is then subjected to a glow discharge treatment for no less than 2 minutes;

[0018] The glow discharge treatment utilizes high-energy particles in the plasma to break chemical bonds on the surface of the PDMS solid matrix, generating active sites that combine with gas molecules such as oxygen and nitrogen to form hydrophilic functional groups;

[0019] At the same time, the etching effect of plasma is used to increase the surface roughness of the PDMS solid substrate.

[0020] Furthermore, the hydrophilic treatment further comprises:

[0021] After the solid substrate is treated with acid, the soaked solid substrate is rinsed with deionized water, and then the solid substrate is soaked in a 0.01%-0.1% (w / v) poly-lysine solution for 5 to 30 minutes to form a hydrophilic covering layer on the surface of the PDMS solid substrate;

[0022] The PDMS solid matrix is ​​then dried.

[0023] Furthermore, the acid used in the acid treatment includes but is not limited to formic acid, acetic acid, hydrochloric acid, lactic acid, and trifluoroacetic acid.

[0024] Furthermore, the film-forming solution is a solution of a single sugar or multiple sugars, and the types of sugars include but are not limited to monosaccharides and polysaccharides.

[0025] Furthermore, the process of releasing the sample from the solid transfer carrier so that the sample meeting the requirements falls off completely from the solid transfer carrier to the target area includes:

[0026] The eluent is used to flush the solid transfer membrane carrying the sample to reduce the adhesion between the solid transfer membrane and the sample, so that the sample can be completely detached from the PDMS solid matrix and flow into the target area together with the eluent to achieve sample recovery.

[0027] Furthermore, the prepared film-forming solution includes:

[0028] Take chitosan raw material, dissolve the chitosan raw material in 1%-5% (v / v) acetic acid solution, and prepare a chitosan solution with a concentration of 0.5%-10% (w / v) as a film-forming solution; or,

[0029] Chitosan and sucrose are prepared by dissolving chitosan in a 1%-5% (v / v) acetic acid solution to prepare a chitosan solution with a concentration of 1-10% (w / v); sucrose is dissolved in deionized water to prepare a sucrose solution with a concentration of 5-20% (w / v); and the two solutions are mixed evenly in a volume ratio of 1:1 to form a film-forming solution;

[0030] The eluent is 0.5%-5% (v / v) acetic acid solution.

[0031] Furthermore, the prepared film-forming solution includes:

[0032] Taking cyclodextrin, preparing the cyclodextrin solution with a concentration of 1%-50% (w / v) as a film-forming solution; or,

[0033] Taking arabinose, preparing the arabinose solution with a concentration of 1%-30% (w / v) as a film-forming solution;

[0034] The eluent is 1× phosphate buffer solution.

[0035] A solid transfer carrier for recovering microdissected samples comprises a PDMS solid substrate, a solid transfer membrane for attaching samples is attached to the PDMS solid substrate, and a hydrophilic treatment layer is provided on the surface of the PDMS solid substrate for enhancing the bonding force between the PDMS solid substrate and the solid transfer membrane.

[0036] Compared with the existing technology, the sample recovery method proposed in this technical solution significantly improves the adsorption recovery rate of the PDMS solid matrix for the cut sample by adding a solid transfer membrane to the PDMS solid matrix. The sample is carried by the solid transfer membrane, which solves the problem of difficulty in transferring the sample from the PDMS solid matrix to the collection tube, and meets the demand for efficient and accurate sample recovery in the field of biological sample research and analysis.

[0037] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention.

[0038] This disclosure is an overview of various implementations or examples of the technology, and is not a comprehensive disclosure of the full scope or all features of the disclosed technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In the drawings, which are not necessarily drawn to scale, the same reference numerals may describe similar components in different views. The same reference numerals with letter suffixes or different letter suffixes may represent different instances of similar components. The accompanying drawings generally illustrate various embodiments by way of example and not limitation, and together with the description and claims, serve to illustrate the embodiments of the invention. Where appropriate, the same reference numerals are used throughout the drawings to refer to the same or similar parts. Such embodiments are illustrative and are not intended to be exhaustive or exclusive of the embodiments of the present apparatus or method.

[0040] Figure 1 Schematic diagram of the solid-state transfer film sample transfer success rate test process according to an embodiment of the present invention.

[0041] Figure 2 Images of sample collection and elution for chitosan-based solid-state transfer membranes.

[0042] Figure 3 Collect samples and post-dissection images for conventional MMI microdissection.

[0043] Figure 4 Schematic diagram of the cutting sample adhesion process. DETAILED DESCRIPTION

[0044] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings of the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the described embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] Unless otherwise defined, the technical or scientific terms used in the present invention shall have the usual meanings understood by persons of ordinary skill in the field to which the present invention belongs. The words "first", "second" and similar terms used in the present invention do not indicate any order, quantity or importance, but are only used to distinguish different components. Words such as "include" or "comprise" mean that the elements or objects preceding the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects. Words such as "connect" or "connected" are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the object being described changes, the relative positional relationship may also change accordingly.

[0046] In order to keep the following description of the embodiments of the present invention clear and concise, detailed descriptions of known functions and known components are omitted.

[0047] See Figures 1 to 3 , an embodiment of the present invention provides a sample recovery method for microdissection, comprising the following steps:

[0048] Making a solid transfer carrier for the sample, wherein the solid transfer carrier at least comprises a PDMS solid matrix and a solid transfer membrane;

[0049] Carrying the sample through the solid transfer membrane so that the sample is attached to the solid transfer carrier;

[0050] When the sample needs to be transferred, the sample is released from the solid transfer carrier so that the sample that meets the needs falls off completely from the solid transfer carrier to the target area.

[0051] This technical solution significantly improves the adsorption recovery rate of the PDMS solid matrix for cut samples by adding a solid transfer membrane to the PDMS solid matrix. The solid transfer membrane carries the sample, solves the problem of difficulty in transferring samples from the PDMS solid matrix to the collection tube, and meets the demand for efficient and accurate sample recovery in the field of biological sample research and analysis.

[0052] In some embodiments, the solid transfer carrier for preparing the sample, comprising at least a PDMS solid matrix and a solid transfer film, includes:

[0053] Prepare a film-forming solution; evenly spread the film-forming solution on the PDMS solid substrate; and dry the film-forming solution to form a solid transfer film on the PDMS solid substrate. Evenly spreading the film-forming solution on the PDMS solid substrate ensures a smooth, even solid transfer film after curing, facilitating sufficient contact with the sample. During use, slowly bring the solid transfer film close to the cut sample and maintain contact for 5 to 10 seconds to ensure sufficient adhesion of the cell sample to the chitosan film.

[0054] In some embodiments, the PDMS solid substrate is pretreated before preparing the solid transfer carrier of the sample, comprising:

[0055] The PDMS solid substrate is subjected to a hydrophilic treatment to enhance the hydrophilicity of the surface of the PDMS solid substrate and improve the bonding strength between the solid transfer membrane and the PDMS solid substrate;

[0056] The hydrophilic treatment includes at least acid treatment;

[0057] The acid treatment comprises:

[0058] The hydrogen ions in the acid act on the silicon-oxygen bonds on the surface of the PDMS solid substrate, partially breaking them and introducing hydroxyl groups to form hydrophilic silicon hydroxyl groups; when the film-forming solution is cured on the PDMS solid substrate, the silicon hydroxyl groups can form hydrogen bonds with water molecules, thereby enhancing the bonding force between the PDMS solid substrate and the solid transfer membrane.

[0059] At the same time, the surface of the PDMS solid substrate is etched with acid to increase the surface roughness of the PDMS solid substrate, thereby enhancing the bonding force between the PDMS solid substrate and the solid transfer membrane; and facilitating the spreading of the film-forming solution on the PDMS solid substrate.

[0060] In some embodiments, the hydrophilic treatment further comprises:

[0061] After the PDMS solid substrate is acid-treated, the PDMS solid substrate is then subjected to a glow discharge treatment for no less than 2 minutes;

[0062] The glow discharge treatment uses high-energy particles in the plasma to break chemical bonds on the surface of the PDMS solid substrate, generating active sites that combine with gas molecules such as oxygen and nitrogen to form hydrophilic functional groups. Specifically, the active sites combine with gas molecules such as oxygen and nitrogen to form hydrophilic functional groups such as hydroxyl, carboxyl, and amino groups.

[0063] At the same time, the etching effect of plasma is used to increase the surface roughness of the PDMS solid substrate.

[0064] In some embodiments, the hydrophilic treatment further comprises:

[0065] After the solid substrate is treated with acid, it is rinsed with deionized water and then immersed in a 0.01%-0.1% (w / v) polylysine solution for 5 to 30 minutes to form a hydrophilic coating on the surface of the PDMS solid substrate. The surface of the PDMS solid substrate is negatively charged, and polylysine contains positively charged amino groups. These positively charged amino groups are electrostatically attracted to the surface of the PDMS solid substrate, and the amino groups can form covalent bonds with the silanol groups on the surface of the PDMS solid substrate. Furthermore, polylysine is a hydrophilic polymer, and the hydrophilic functional groups on its molecular chain form a hydrophilic coating on the surface of the PDMS solid substrate, thereby increasing the hydrophilicity of the PDMS solid substrate.

[0066] The PDMS solid matrix is ​​then dried.

[0067] In some embodiments, the acid used in the acid treatment includes, but is not limited to, formic acid, acetic acid, hydrochloric acid, lactic acid, and trifluoroacetic acid. When formic acid is used for the acid treatment, the concentration of the formic acid solution can range from 0.5% to 100% (v / v); when hydrochloric acid is used, the concentration of the hydrochloric acid solution can range from 0.5% to 32% (v / v). For other acids, such as acetic acid, lactic acid, and trifluoroacetic acid, the concentration can be adjusted within a reasonable range based on actual experimental results, the sample, and the characteristics of the PDMS solid matrix.

[0068] The treatment time ranges from 0.05 to 12 hours and needs to be adjusted according to the acid concentration. High-concentration acids have high reactivity and require shorter treatment times. For example, for formic acid with a concentration of over 50% or hydrochloric acid with a concentration of over 20%, the treatment time may be controlled at 0.05 to 1 hour to avoid damage to the sample and solid matrix caused by excessive treatment. Low-concentration acids react slowly and require longer treatment times. For example, for formic acid or hydrochloric acid with a concentration of less than 5%, the treatment time may be 1 to 12 hours to ensure the desired hydrophilic treatment effect. At the same time, the characteristics of the sample and solid matrix must be taken into consideration. For acid-sensitive samples, the treatment time should be appropriately shortened even when using low-concentration acids; for samples and solid matrices with strong tolerance, the treatment time can be appropriately extended.

[0069] In some embodiments, the film-forming solution is a solution of a single sugar or multiple sugars, and the types of sugars include but are not limited to monosaccharides and polysaccharides. For example: chitosan, cyclodextrin, arabinose, sucrose, etc. The solid transfer membrane prepared from the film-forming solution using sugar as raw material has high adhesion, which can make the sample more effectively adhere to the PDMS solid matrix during the collection process, greatly reducing sample loss. The solid transfer membrane prepared using a film-forming solution using a single sugar as raw material, such as a film-forming solution using chitosan as raw material, can have a sample recovery rate of up to 90%, or even higher; using a film-forming solution mixed with multiple sugars can also achieve a fairly high recovery rate. Carbohydrate materials are low-cost and easy to obtain and prepare, which makes the entire solution practical and economical, and suitable for wide promotion and application in scientific research, clinical diagnosis and other fields.

[0070] In some embodiments, the step of releasing the sample from the solid transfer carrier so that the sample meeting the requirements falls completely off the solid transfer carrier to the target area includes:

[0071] The eluent is used to flush the solid transfer membrane carrying the sample, reducing the adhesion between the solid transfer membrane and the sample, allowing the sample to completely fall off the PDMS solid substrate and flow into the target area along with the eluent, achieving sample recovery. Specifically, after the eluent comes into contact with the solid transfer membrane, the solid transfer membrane dissolves in the eluent, transforming from a solid to a liquid state. As the dissolution process proceeds, the sample adsorbed on the solid transfer membrane falls off and enters the eluent, where it can flow into the target area (such as a collection tube) along with the eluent.

[0072] In some embodiments, the prepared film-forming solution comprises:

[0073] Taking chitosan raw materials, dissolving the chitosan raw materials in 1%-5% (v / v) acetic acid solution to prepare a chitosan solution with a concentration of 0.5%-10% (w / v) as a film-forming solution;

[0074] or,

[0075] Chitosan and sucrose are prepared by dissolving chitosan in a 1%-5% (v / v) acetic acid solution to prepare a chitosan solution with a concentration of 1-10% (w / v); sucrose is dissolved in deionized water to prepare a sucrose solution with a concentration of 5-20% (w / v); and the two solutions are mixed evenly in a volume ratio of 1:1 to form a film-forming solution;

[0076] The eluent is 0.5%-5% (v / v) acetic acid solution.

[0077] In some embodiments, the prepared film-forming solution comprises:

[0078] Taking cyclodextrin, preparing the cyclodextrin solution with a concentration of 1%-50% (w / v) as a film-forming solution; or,

[0079] Taking arabinose, preparing the arabinose solution with a concentration of 1%-30% (w / v) as a film-forming solution;

[0080] The eluent is 1× phosphate buffer solution.

[0081] Test on the transfer success rate of solid-state transfer membrane samples prepared from membrane-forming solutions containing different types of sugars:

[0082] Preparation of solid-state transfer membranes

[0083] 1. Solid transfer membrane prepared from a film-forming solution with chitosan as raw material: Prepare chitosan raw material and dissolve it in 2% (v / v) acetic acid solution (acetic acid solution fully protonates chitosan and dissolves chitosan, which forms a strong electrostatic bond with the cell membrane phospholipid bilayer (negatively charged)). Prepare a chitosan solution with a concentration of 3% (w / v) (chitosan molecular chains can form a monolayer network on the surface of the PDMS solid matrix, with a moderate amino density, which can capture negatively charged cells (such as sialic acid on the surface of tumor cells) through electrostatic action, and avoid difficulty in dissolving during elution due to the dense entanglement of molecular chains. After Add 10 μl of chitosan solution to a pretreated PDMS solid substrate (such as a PDMS-PCR tube cap or MMI tube cap, including conventional MMI tube caps and various adapter tube caps), ensuring the chitosan solution is concentrated in the center of the PDMS solid substrate. Allow to air dry at room temperature or in an oven at 30-40°C to form a uniform solid transfer film of chitosan on the surface of the PDMS solid substrate. Avoid high temperatures during drying, as these can cause thermal oxidation of the chitosan amino groups. A temperature of 30-40°C maintains the adhesion of the resulting solid transfer film while allowing the solution's water to evaporate quickly to form a film.

[0084] 2. Solid-state transfer membrane prepared from a film-forming solution using cyclodextrin as the raw material: Prepare a cyclodextrin solution with a concentration of 5% (w / v). A cyclodextrin solution with a concentration of 1%-50% has good solubility in pure water, allowing for uniform film formation without overly dense film layers due to high concentrations, which could affect the entry and release of small molecules. Then, similar to the chitosan membrane, add 10 μl of cyclodextrin solution to the pretreated tube cap. Allow to dry naturally at room temperature or in an oven at 30-40°C to allow the cyclodextrin to form a uniform solid transfer membrane on the surface of the PDMS solid substrate.

[0085] 3. Solid-state transfer membrane prepared with a film-forming solution containing arabinose: Prepare a 10% (w / v) arabinose solution. (Concentrations below 1% result in insufficient film viscosity, making it prone to slippage, especially on wet samples. Concentrations above 30% are too viscous, making it difficult to spread evenly and prone to cracking.) Then, similar to the chitosan film, add 10 μl of the arabinose solution to the pretreated tube cap. Allow to air dry at room temperature or dry in an oven at 30-40°C to form a uniform solid transfer membrane of arabinose on the surface of the PDMS substrate.

[0086] 4. Solid-state transfer membrane prepared from a film-forming solution containing multiple carbohydrates (using chitosan and sucrose as an example): Prepare chitosan and sucrose separately. Dissolve chitosan in 2% (v / v) acetic acid to create a 6% (w / v) chitosan solution. Dissolve sucrose in deionized water to create a 10% (w / v) sucrose solution. Mix the two solutions in a 1:1 volume ratio. Add 10 µl of the mixed solution to the cap of the pretreated tube and allow to air dry at room temperature or in an oven at 30-40°C to form a solid-state transfer membrane containing a complex carbohydrate bonded together with chitosan and sucrose. The amino groups of chitosan provide electrostatic adsorption, while the multiple hydroxyl groups of sucrose enhance hydrogen bonding. The combination of the two forms a "double network" adhesion layer, which has strong capture capabilities for samples of varying charge properties.

[0087] Sample recovery experiment

[0088] Use MMI laser microcutting equipment to cut the sample slices and place the cut samples on the laboratory table.

[0089] A PDMS solid substrate containing a solid transfer membrane made from a membrane-forming solution containing chitosan was slowly brought close to the sample after slicing. Contact was maintained for 5-10 seconds to allow the cell sample to adhere to the membrane. The eluent used was a solution containing 0.5% (v / v) acetic acid. (A 0.5%-5% (v / v) acetic acid solution has a pH and ionic strength close to that of the intracellular environment, maintaining the cell's state. This solution dissolves the carbohydrate-based solid transfer membrane from a solid state to a liquid state, allowing the sample adsorbed on the membrane to detach and disperse in the eluent. This gentle process avoids damage to the sample.) An appropriate amount of this eluent was added to the PDMS solid substrate and gently agitated. Following the interaction of the eluent with the chitosan, the cell sample gradually detached from the PDMS solid substrate and flowed out with the eluent, completing sample collection. The collected sample was evaluated microscopically. Results showed a sample recovery rate exceeding 90%. Figure 2 The image shows the sample collected by the solid transfer membrane using chitosan as raw material and eluted with 0.5% (v / v) acetic acid solution. Figure 3 The images show samples collected using conventional MMI microdissection and eluted with 1× phosphate buffer solution. A comparison shows that the sample collected using the chitosan membrane was completely transferred to the eluent after elution. However, the sample collected using conventional MMI microdissection was only partially transferred after elution, with residual sample remaining on the tube cap. This significantly lower sample transfer efficiency than the chitosan membrane. This demonstrates that the chitosan membrane performs better in sample transfer.

[0090] For the PDMS solid substrate with a solid transfer membrane prepared with a membrane-forming solution using cyclodextrin as the raw material, the above-mentioned cut sample adhesion process was also carried out, and 1X phosphate buffer solution (PBS) was selected as the eluent. (1× phosphate buffer solution is used as the eluent. Its pH value and ionic strength are close to the intracellular environment and can maintain the cell state. It can dissolve the solid transfer membrane of the carbohydrate raw material from solid to liquid, allowing the sample adsorbed on the membrane to detach and disperse in the eluent. The whole process is gentle and avoids damage to the sample.) Experimental results show that the sample transfer success rate is about 90%.

[0091] For a PDMS solid-state transfer membrane prepared with a membrane-forming solution containing arabinose, the same cut-sample adhesion process was repeated, followed by elution using 1X phosphate buffered saline (PBS). Preliminary experiments showed that under optimized conditions, sample recovery and transfer were also effective, with a sample transfer success rate of approximately 90%.

[0092] The same process for adhering the cut sample to a PDMS substrate with a solid transfer membrane prepared from a membrane-forming solution containing chitosan and sucrose was repeated. Using 0.5% (v / v) acetic acid as the eluent effectively reduced the adhesion. Evaluation by relevant testing methods demonstrated sample recovery rates exceeding 95%, demonstrating excellent sample quality.

[0093] See Figure 4 An embodiment of the present invention provides a solid-state transfer medium for recovering microdissected samples, comprising a PDMS solid substrate 1, attached to which is a solid-state transfer membrane 2 for attaching the sample. The surface of the PDMS solid substrate 1 is provided with a hydrophilic treatment layer, which is used to enhance the bonding between the PDMS solid substrate 1 and the solid-state transfer membrane 2. The solid-state transfer medium proposed in this technical solution does not require large-scale modification of existing microdissection equipment (including but not limited to MMI laser microdissection and Leica laser microdissection equipment).

[0094] The above description is intended to be illustrative rather than restrictive, and those skilled in the art may make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present disclosure. Furthermore, the above examples (or one or more of them) may be used in combination with each other, and it is contemplated that these embodiments may be combined with each other in various combinations or permutations.

Claims

1. A method for recovering a sample for microdissection, characterized in that: The steps include: Making a solid transfer carrier for the sample, wherein the solid transfer carrier at least comprises a PDMS solid matrix and a solid transfer membrane; Carrying the sample through the solid transfer membrane so that the sample is attached to the solid transfer carrier; When the sample needs to be transferred, the sample is released from the solid transfer carrier so that the sample that meets the needs falls off completely from the solid transfer carrier to the target area.

2. The sample recovery method for microdissection according to claim 1, characterized in that: The solid transfer carrier for preparing the sample, which at least includes a PDMS solid matrix and a solid transfer film, includes: A film-forming solution is prepared; the film-forming solution is evenly spread on the PDMS solid substrate; and the film-forming solution is dried to form a solid transfer film on the PDMS solid substrate.

3. The sample recovery method for microdissection according to claim 1, characterized in that: The PDMS solid matrix is ​​pretreated before preparing the solid transfer carrier of the sample, including: The PDMS solid substrate is subjected to a hydrophilic treatment to enhance the hydrophilicity of the surface of the PDMS solid substrate and improve the bonding strength between the solid transfer membrane and the PDMS solid substrate; The hydrophilic treatment includes at least acid treatment; The acid treatment comprises: The hydrogen ions in the acid react with the silicon-oxygen bonds on the surface of the PDMS solid substrate, partially breaking them and introducing hydroxyl groups to form hydrophilic silanol groups; At the same time, the surface of the PDMS solid substrate is etched with acid to increase the surface roughness of the PDMS solid substrate.

4. The sample recovery method for microdissection according to claim 3, characterized in that: The hydrophilic treatment further comprises: After the PDMS solid substrate is acid-treated, the PDMS solid substrate is then subjected to a glow discharge treatment for no less than 2 minutes; The glow discharge treatment utilizes high-energy particles in the plasma to break chemical bonds on the surface of the PDMS solid matrix, generating active sites that combine with gas molecules such as oxygen and nitrogen to form hydrophilic functional groups; At the same time, the etching effect of plasma is used to increase the surface roughness of the PDMS solid substrate.

5. The sample recovery method for microdissection according to claim 3, characterized in that: The hydrophilic treatment further comprises: After the solid substrate is treated with acid, the soaked solid substrate is rinsed with deionized water, and then the solid substrate is soaked in a 0.01%-0.1% (w / v) poly-lysine solution for 5 to 30 minutes to form a hydrophilic covering layer on the surface of the PDMS solid substrate; The PDMS solid matrix is ​​then dried.

6. The sample recovery method for microdissection according to claim 3, characterized in that: The acid used in the acid treatment includes but is not limited to formic acid, acetic acid, hydrochloric acid, lactic acid, and trifluoroacetic acid.

7. The sample recovery method for microdissection according to claim 2, characterized in that: The film-forming solution is a solution of a single sugar or multiple sugars, and the types of sugars include but are not limited to monosaccharides and polysaccharides.

8. The sample recovery method for microdissection according to claim 2 or 7, characterized in that: The method of releasing the sample from the solid transfer carrier so that the sample meeting the requirements falls off from the solid transfer carrier completely to the target area includes: The eluent is used to flush the solid transfer membrane carrying the sample to reduce the adhesion between the solid transfer membrane and the sample, so that the sample can be completely detached from the PDMS solid matrix and flow into the target area together with the eluent to achieve sample recovery.

9. The sample recovery method for microdissection according to claim 8, characterized in that: The prepared film-forming solution comprises: Take chitosan raw material, dissolve the chitosan raw material in 1%-5% (v / v) acetic acid solution, and prepare a chitosan solution with a concentration of 0.5%-10% (w / v) as a film-forming solution; or, Chitosan and sucrose are prepared by dissolving chitosan in a 1%-5% (v / v) acetic acid solution to prepare a chitosan solution with a concentration of 1-10% (w / v); sucrose is dissolved in deionized water to prepare a sucrose solution with a concentration of 5-20% (w / v); and the two solutions are mixed evenly in a volume ratio of 1:1 to form a film-forming solution; The eluent is 0.5%-5% (v / v) acetic acid solution.

10. The sample recovery method for microdissection according to claim 8, characterized in that: The prepared film-forming solution comprises: Taking cyclodextrin, preparing the cyclodextrin solution with a concentration of 1%-50% (w / v) as a film-forming solution; or, Taking arabinose, preparing the arabinose solution with a concentration of 1%-30% (w / v) as a film-forming solution; The eluent is 1× phosphate buffer solution.

11. A solid transfer carrier for recovering microdissected samples, comprising a PDMS solid matrix, characterized in that: A solid transfer membrane for attaching a sample is attached to the PDMS solid substrate, and a hydrophilic treatment layer is provided on the surface of the PDMS solid substrate to enhance the bonding force between the PDMS solid substrate and the solid transfer membrane.

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