Paraffin section pasting method capable of being used for space transcriptomics and application

Abstract

The invention discloses a paraffin section pasting method capable of being used for space transcriptomics and application, and belongs to the technical field of paraffin sections.The paraffin section pasting method comprises the following steps that sample paraffin blocks needing to be researched are cut into paraffin rolls; a round knife is used for buckling and pressing the wax roll, and a target tissue area is cut; inflating the tail part of the circular knife through an injector, and blowing out a cut complete target tissue piece; adsorbing the complete target tissue piece by using a needle head with static electricity; and heating the space transcription chip, dropwise adding ultrapure water into a target patch area on the space transcription chip to form liquid drops, enabling a needle head with static electricity adsorbed with the complete target tissue slice to approach the target patch area, and attracting the complete target tissue slice onto the liquid drops under the electrostatic action to complete paraffin section patch. According to the method, the wax roll is used for splicing a plurality of samples, damage to the wax block is avoided, the manufacturing process is simple, high technical level and professional equipment are not needed, and therefore the manufacturing cost is low.

Description

Technical Field

[0001] The present invention relates to the technical field of paraffin sections, and in particular to a paraffin section patching method and application that can be used for spatial transcriptomics. Background Art

[0002] Spatial transcriptomics is an emerging technology that simultaneously obtains gene expression signatures and spatial distribution data in situ in tissues, revealing the spatial heterogeneity of gene expression and providing an important tool for understanding complex biological processes and disease mechanisms.

[0003] While spatial transcriptomics is a powerful technology, it also has drawbacks and limitations. The most prominent of these is the relatively high cost of sequencing, which can cost as much as 40,000 to 70,000 RMB per sample, limiting its widespread application in clinical and scientific research.

[0004] Therefore, developing new technologies to enable the detection of multiple samples on the same chip and improve the cost-effectiveness of spatial transcriptomics chips is an urgent problem to be solved.

[0005] Existing tissue microarray technology enables the analysis of multiple samples on a single spatial transcriptomics chip. Scientists extract tissue from conventional tissue blocks and rearrange these tissues into an array, allowing multiple cases or tissue types to be integrated into a single slide. This tissue microarray allows researchers to obtain high-throughput gene expression information from the same slide.

[0006] However, tissue chips require drilling and sampling from existing embedded samples and reassembling them into chip wax blocks, which is bound to damage the original wax blocks. However, in clinical practice, pathological wax blocks have important preservation value and must not be subjected to destructive research. In addition, the production instruments and analysis systems for tissue chips are currently expensive and have not yet been widely promoted and applied. Tissue chips generally require clear experimental purposes and fixed groups because the chip wax blocks are inconvenient to change after being wrapped. In actual research situations, sometimes it is necessary to reassemble or only study individual samples, and the array embedding blocks are very inflexible.

[0007] Therefore, it is of great significance to develop a non-destructive, flexible and economical "tissue chip" technology. Summary of the Invention

[0008] The present invention provides a paraffin section patch method and application that can be used for spatial transcriptomics. The sample organization of the present invention is flexible, and only wax rolls of related samples are needed, which can be combined at will, and has high flexibility.

[0009] The present invention solves its technical problems by adopting the following technical solutions: A paraffin section mounting method for spatial transcriptomics comprises the following steps: Cut the sample wax block to be studied into wax rolls; Use a circular knife to press on the wax roll and cut out the target tissue area; Inflate the circular knife tail with a syringe to blow out the complete target tissue slice; The complete target tissue piece is adsorbed with an electrostatically charged needle; The spatial transcription chip is heated, and ultrapure water is added to the target patch area on the spatial transcription chip to form droplets. The electrostatically charged needle with the complete target tissue piece adsorbed is brought close to the target patch area. Under the action of static electricity, the complete target tissue piece is attracted to the droplet, completing the paraffin section patch.

[0010] As a preferred embodiment of the present invention, the thickness of the wax roll is 4-6 μm.

[0011] As a preferred embodiment of the present invention, the diameter of the circular knife is 2-4 mm.

[0012] As a preferred embodiment of the present invention, the syringe is without the needle.

[0013] As a preferred embodiment of the present invention, the heating temperature is 40-50°C.

[0014] As a preferred embodiment of the present invention, the heating temperature is 45°C.

[0015] As a preferred embodiment of the present invention, the amount of ultrapure water added is 2 to 5 μL.

[0016] As a preferred embodiment of the present invention, static electricity is generated by rapidly rubbing the syringe barrel on a latex glove, thereby causing the needle to be electrostatically charged.

[0017] The present invention also provides an application of paraffin sections in spatial transcriptomics chip detection, wherein the paraffin sections adopt the paraffin section mounting method described above.

[0018] The beneficial effects of the present invention are as follows: the present invention uses wax rolls to piece together multiple samples, thus avoiding damage to the wax blocks; the production process of the present invention is simple, does not require a high level of technology and professional equipment, and therefore has a low manufacturing cost; the sample organization of the present invention is flexible, and only wax rolls of related samples are required, which can be combined at will, and has high flexibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a picture of using a 3mm diameter round knife to press the wax roll.

[0020] Figure 2 Preheat the spatial transcriptome microarray on the heating stage.

[0021] Figure 3The prepared paraffin section patch. DETAILED DESCRIPTION

[0022] To make the purpose, technical solutions, and advantages of the embodiments of this application more clear, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of this application, not all of them. Based on the embodiments of this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0023] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0024] In this application, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0025] In the present application, there is no particular limitation on the specific dispersion and stirring treatment methods.

[0026] Unless otherwise specified, the components, raw materials or instruments used in the embodiments and comparative examples of the present invention are all commercially available raw materials or instruments, and the components and raw materials used in each parallel experiment are all of the same kind.

[0027] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "plurality" means at least two, such as two, three, etc., unless otherwise specifically defined.

[0028] Example 1 A paraffin section mounting method for spatial transcriptomics comprises the following steps: (1) Prepare materials: 3mm diameter circular knife, tweezers, 5ml syringe, insulin needle, constant temperature heating table.

[0029] (2) Use a microtome to cut the wax block of the sample to be studied into 5 μm thick wax rolls.

[0030] (3) If Figure 1As shown, a 3 mm diameter circular knife was used to press on the wax roll to cut out the target tissue area.

[0031] (4) Use a 5 ml syringe (without the needle) to quickly pump air into the tail of the circular knife to blow out the complete circular tissue piece.

[0032] (5) Quickly rub the insulin syringe on a latex glove to generate static electricity, then bring the insulin needle close to the circular tissue piece and use static electricity to attract the entire circular tissue piece to the needle.

[0033] (6) If Figure 2 As shown, preheat the spatial transcription chip to 45 degrees on a heating table, then add 3.5ul of ultrapure water to the target patch area on the spatial transcription chip, and carefully move the insulin needle with the tissue piece close to the droplet. Under the action of static electricity, the tissue piece will be attracted to the droplet.

[0034] (7) Repeat the above steps to apply the next target sample. Continue heating at 45 degrees for 2 hours to spread and dry the sample.

[0035] Finally, it should be noted that the above embodiments are intended to illustrate the technical solutions of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A paraffin section patch method for spatial transcriptomics, characterized in that: The following steps are involved: Cut the wax block of the sample to be studied into wax rolls; Use a circular knife to press on the wax roll and cut out the target tissue area; Inflate the circular knife tail with a syringe to blow out the complete target tissue slice; The complete target tissue piece is adsorbed with an electrostatically charged needle; The spatial transcription chip is heated, and ultrapure water is added to the target patch area on the spatial transcription chip to form droplets. The electrostatically charged needle with the complete target tissue piece adsorbed is brought close to the target patch area. Under the action of static electricity, the complete target tissue piece is attracted to the droplet, completing the paraffin section patch.

2. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The thickness of the wax roll is 4-6 μm.

3. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The diameter of the circular knife is 2-4 mm.

4. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The syringe has the needle removed.

5. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The heating temperature is 40-50°C.

6. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The heating temperature is 45°C.

7. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The amount of ultrapure water added is 2-5 μL.

8. The paraffin section mounting method for spatial transcriptomics according to claim 1, characterized in that: The needle is charged with static electricity by rubbing the syringe barrel rapidly against a latex glove to generate static electricity.

9. An application of paraffin sections in spatial transcriptomics chip detection, characterized in that: The paraffin section adopts the paraffin section pasting method according to any one of claims 1 to 8.

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