A knife and collection system for laser microdissection

Abstract

This application discloses a cutting tool and collection system for laser micro-cutting. The cutting tool includes a sample stage, a tool body, a fan, and a microscope objective. The tool body has an airflow channel that runs through it, with a first air inlet and a first air outlet at each end. The cutting head is located at the first air inlet, and the first air outlet is connected to the fan. Thus, the fan draws in gas and creates a negative pressure area at the cutting head. The sample stage moves the tissue block closer to the cutting head, causing the cutting head to cut the tissue block and form tissue sections. The tissue sections remain connected to the tissue block and are not completely separated from it. Under controllable negative pressure, the tissue sections adhere tightly to the cutting head, thereby significantly reducing tissue section displacement, warping, and detachment caused by cutting vibration, fluid disturbance, or laser scanning. This improves the spatial stability and positioning accuracy of the tissue sections during laser micro-cutting.

Description

Technical Field

[0001] This application relates to the field of microscopic section preparation and histological research technology, specifically to a cutting tool and collection system for laser micro-cutting. Background Technology

[0002] Acquiring spatial information at the single-cell level is a key technological requirement in current biomedical research, spatial transcriptomics, tumor heterogeneity analysis, and neuroscience research. To achieve precise acquisition of specific cells or cell populations in complex biological tissues, researchers typically need to perform microscale separation and retrieval of target regions while preserving as much of the original spatial structure information of the tissue as possible.

[0003] Laser capture microdissection (LCM) technology is widely used for separating target micro-regions in tissue sections due to its advantages of non-contact operation and high precision. However, existing laser microdissection techniques typically use pre-prepared two-dimensional tissue sections attached to a glass slide as the operating object. This presents limitations in sample preparation, preservation of spatial information, and cutting efficiency, especially when directly obtaining single cells or cell populations from tissue blocks, making it difficult to achieve an efficient, continuous, and low-contamination workflow. Summary of the Invention

[0004] This application aims to address at least one of the technical problems existing in the prior art. To this end, this application proposes a cutting tool for laser microdissection, which can stably fix tissue sections during the cutting process, prevent contamination during the collection of tissue sections, and achieve efficient recovery.

[0005] This application also proposes a collection system for laser micro-cutting.

[0006] A cutting tool for laser microsurgery according to one embodiment of this application includes: a sample stage, which is movable, with a tissue block fixed on one side of the sample stage in a first direction, and the movement of the sample stage can drive the tissue block to move; a cutting tool body, which is fixedly disposed in the first direction of the sample stage; the cutting tool body includes a cutting head and an airflow channel, the airflow channel penetrating the cutting tool body and forming a first air inlet and a first air outlet on the surface of the cutting tool body, the cutting head being used to cut the tissue block to form tissue sections, the cutting head being disposed at the port of the first air inlet, the cutting head being located outside the first air inlet and inclined relative to the sample stage; an exhaust fan, the exhaust fan being connected to the first air outlet of the cutting tool body to extract gas; and a microscope objective, the microscope objective being disposed on one side of the cutting tool body, the microscope objective being used to conduct ultraviolet laser cutting light path to micro-cut tissue sections to form single cells or cell groups.

[0007] According to one embodiment of this application, the blade head includes a blade surface located on one side of the blade head in a first direction. The sample stage can drive the tissue block to move and contact the blade head. The blade head cuts the tissue block to form a tissue section, and the tissue section is located on the blade surface.

[0008] According to one embodiment of this application, the first air inlet is located in a first direction of the cutter head.

[0009] According to one embodiment of this application, the microscope objective is disposed opposite to the tool body, the lens of the microscope objective is directly facing the cutting surface of the tool body, and both the microscope objective and the tool body are disposed in a first direction on the sample stage.

[0010] According to one embodiment of this application, a microscope objective is used for real-time microscopic observation of tissue sections and for imaging and positioning tissue sections before microscopic cutting of tissue sections.

[0011] A collection system for laser micro-cutting according to one embodiment of this application includes: the aforementioned cutting tool for laser micro-cutting; a gas guiding assembly, which has a gas guiding cavity, the gas guiding assembly including a first end and a second end, the gas guiding cavity connecting the first end and the second end, the first end being connected to a first air outlet of the cutting tool body, and the second end being connected to an exhaust fan, the exhaust fan being used to extract gas sequentially through the airflow channels of the gas guiding assembly and the cutting tool body; and a cell sieve, which is disposed in the gas guiding assembly and separates the gas guiding cavity to screen target single cells or target cell populations.

[0012] According to another embodiment of this application, the cell sieve is provided with two or more layers, and the cell sieves are arranged sequentially along the airflow direction in the air guide cavity in descending order of sieve aperture size.

[0013] According to another embodiment of this application, a cell sieve filter is also included. The cell sieve filter includes a receiving member with a receiving cavity inside. The cell sieve filter has a second air inlet on its outer wall, which communicates with the receiving cavity. The cell sieve filter also includes a blocking member that extends into the receiving cavity. One end of the blocking member that extends into the receiving cavity has a second air outlet that penetrates the blocking member and communicates with the outer wall of the cell sieve filter. The first air outlet of the blade body is connected to the second air inlet, and the exhaust fan is connected to the second air outlet.

[0014] According to another embodiment of this application, the cell sieve filter further includes a filter element having multiple micropores, the filter element being disposed at a second air outlet, the micropores being capable of blocking cells and cell clusters.

[0015] According to another embodiment of this application, the air guiding assembly includes a first air guiding pipe and a second air guiding pipe; one end of the first air guiding pipe serves as the first end of the air guiding assembly and is connected to a first air outlet, and the other end is connected to a second air inlet; one end of the second air guiding pipe serves as the second end of the air guiding assembly and is connected to an exhaust fan, and the other end is connected to a second air outlet.

[0016] The tissue section cutting and collecting device according to the embodiments of this application has at least the following beneficial effects: the blade body is provided with an airflow channel that runs through the blade body, and the two ends of the airflow channel are a first air inlet and a first air outlet, respectively. The blade head is located at the first air inlet port, and the first air outlet is connected to an exhaust fan. Thus, the exhaust fan draws in gas and forms a negative pressure area at the blade head. The sample stage drives the tissue block to move close to the blade head and causes the blade head to cut the tissue block to form tissue sections. The formed tissue sections are always connected to the tissue block and are not completely separated from the tissue block. Under the controllable negative pressure, the tissue sections are tightly attached to the blade head, thereby significantly reducing the displacement, warping and detachment of tissue sections caused by cutting vibration, fluid disturbance or laser scanning, and improving the spatial stability and positioning accuracy of tissue sections in the laser micro-cutting process.

[0017] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0018] The present application will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 This is a schematic diagram of the collection system for laser micro-cutting according to an embodiment of this application; Figure 2 for Figure 1 Side view of the collection system used for laser micro-cutting; Figure 3 for Figure 2 Enlarged schematic diagram of section A of the structure; Figure 4 This is a cross-sectional view of the tool body in a tool for laser micro-cutting according to another embodiment of this application; Figure 5 for Figure 1 A cross-sectional view of the first gas duct in a collection system for laser micro-cutting; Figure 6 for Figure 1 A cross-sectional view of a cell sieve filter in a collection system used for laser microdissection; Figure 7 A cross-sectional view of a cell sieve filter with baffles.

[0019] Figure label: 10. Tissue block; 11. Tissue section; 100 Sample stage; 200 Tool body; 210 Tool head; 211 Tool face; 220 First air inlet; 230 First air outlet; 300 Exhaust fan; 400 Microscope objective; 410 Dropper; 500 Gas guiding assembly; 501 First end; 502 Second end; 503 Gas guiding chamber; 510 First gas guiding tube; 520 Second gas guiding tube; 600 Cell sieve; 700 Cell sieve filter; 710 Receptacle; 711 Receptacle; 720 Second air inlet; 730 Blocking element; 731 Second air outlet; 740 Filter element; 750 Baffle; 800 Support. Detailed Implementation

[0020] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0021] In the description of this application, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0022] In the description of this application, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0023] In the description of this application, unless otherwise expressly defined, terms such as "setup," "installation," and "connection" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this application in conjunction with the specific content of the technical solution.

[0024] In the description of this application, the terms "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0025] Acquiring spatial information at the single-cell level is a key technological requirement in current biomedical research, spatial transcriptomics, tumor heterogeneity analysis, and neuroscience research. To achieve precise acquisition of specific cells or cell populations in complex biological tissues, researchers typically need to perform microscale separation and retrieval of target regions while preserving as much of the original spatial structure information of the tissue as possible.

[0026] Laser capture microdissection (LCM) technology, with its advantages of non-contact operation and high-precision positioning, has been widely used for the separation and acquisition of target micro-regions in tissue sections. However, existing laser microdissection techniques typically use pre-prepared two-dimensional tissue sections attached to glass slides as the operating objects, which still has certain limitations in terms of sample preparation process, spatial information preservation, and cutting efficiency. Especially for applications that directly obtain single cells or cell populations from tissue blocks, existing technologies struggle to achieve efficient, continuous, and low-contamination operating procedures.

[0027] One embodiment of this application discloses a cutting tool for laser micro-cutting; please refer to [link to relevant documentation]. Figure 1 and Figure 2 It includes a sample stage 100, a microscope objective 400, and a cutting tool body 200. The top surface of the sample stage 100 serves as a loading surface for fixing and placing the tissue block 10. The cutting tool body 200 is fixedly mounted on the instrument's support 800, located above the loading surface. The cutting tool body 200 includes a cutting head 210 for cutting. It is understood that... Figure 2The direction indicated by the arrow on the X-axis is above the sample stage 100. The sample stage 100 is a three-axis precision displacement stage, which allows the sample stage 100 to move the tissue block 10 close to the cutter head 210 of the cutter body 200. The tissue block 10 moves relative to the cutter body 200, and the cutter head 210 of the cutter body 200 cuts the tissue block 10 to form a tissue section 11. The microscope objective 400 is also positioned above the sample stage 100 and opposite to the cutter body 200 to avoid interference between the two. The microscope objective 400 can transmit the ultraviolet laser cutting light path to the tissue section 11 to perform micro-cutting on the target micro-regions in the tissue section 11, thereby obtaining target single cells or target cell populations. In addition, the cutter head 210 is tilted at a certain angle relative to the loading surface of the sample stage 100 so that the cutter head 210 can cut the tissue block 10 with better cutting effect. The specific angle of tilt can be determined according to actual needs, and therefore is not limited here.

[0028] Further, please see Figure 2 , Figure 3 and Figure 4 The tool body 200 of the laser micro-cutting tool of this application has an internal airflow channel. The two ends of the airflow channel penetrate the surface of the tool body 200 and form a first air inlet 220 and a first air outlet 230, respectively. The cutting head 210 of the tool body 200 is located at the port of the first air inlet 220. The laser micro-cutting tool also includes a fan 300. The exhaust end of the fan 300 is connected to the first air outlet 230 of the tool body 200. Thus, the fan 300 can extract gas and form a negative pressure area at the cutting head 210.

[0029] Understandably, the exhaust fan 300 remains on throughout the process of the cutter body 200 cutting the tissue block 10 and the microscope objective 400 transmitting ultraviolet laser light to micro-cut the tissue section 11. The sample stage 100 moves the tissue block 10 closer to the cutter head 210, causing the cutter head 210 to cut the tissue block 10 to form the tissue section 11. The formed tissue section 11 is always connected to the tissue block 10 and is not completely separated from the tissue block 10. Under the controllable negative pressure of the exhaust fan 300, the tissue section 11 is tightly attached to the cutter head 210. As a result, in the subsequent ultraviolet laser micro-cutting process of the tissue section 11, the displacement, warping and detachment of the tissue section 11 caused by micro-cutting vibration, fluid disturbance or laser scanning are significantly reduced, and the spatial stability and positioning accuracy of the tissue section 11 during the laser micro-cutting process are improved.

[0030] Further, please see Figure 3The cutting head 210 includes a cutting surface 211, which is located on one side of the cutting head 210 in a first direction. The sample stage 100 can move the tissue block 10 to contact the cutting head 210. The cutting head 210 cuts the tissue block 10 to form a tissue section 11, which is located on the cutting surface 211. That is, the cutting head 210 has a flat cutting surface 211 on its upper side. The sample stage 100 moves the tissue block 10 gradually closer to the lower end of the cutting head 210 in a horizontal direction. The cutting head 210 cuts the tissue block 10, and the resulting tissue section 11 is raised upwards. However, under the action of a negative pressure area formed at the first air inlet 220 of the cutting tool body 200, such as... Figure 3 As indicated by the arrow, the gas flows towards the first air inlet 220 in the direction of the arrow. The tissue slice 11 tends to move towards the first air inlet 220 under the influence of the airflow. Since the tissue slice 11 is not completely separated from the tissue block 10, the tissue slice 11 can only adhere to the blade surface 211 on the upper side of the blade head 210. The upper end of the tissue slice 11 is pulled by the negative pressure, and the lower end is always connected to the tissue block 10. Thus, the tissue slice 11 remains taut, avoiding displacement and warping, and also improving the stability and positioning accuracy when micro-cutting it later.

[0031] Furthermore, such as Figure 3 As shown, the first air inlet 220 is located above the blade head 210. This means that the negative pressure area formed at the first air inlet 220 is above the blade head 210, and the traction direction of the negative pressure area on the tissue slice 11 is obliquely upward. In this case, the traction effect is better, which helps to keep the tissue slice 11 taut. Of course, the position of the first air inlet 220 relative to the blade head 210 can also be below it. In this case, the negative pressure area formed at the first air inlet 220 can also traction the upper end of the tissue slice 11. Compared with the aforementioned first air inlet 220 being above the blade head 210, the only difference is in the traction effect. It can be understood that when the first air inlet 220 is above the blade head 210, the negative pressure area has a better traction effect on the tissue slice 11.

[0032] In some embodiments, the microscope objective 400 is disposed opposite to the tool body 200, with the lens of the microscope objective 400 facing the cutting surface 211 of the tool body 200. Both the microscope objective 400 and the tool body 200 are disposed in a first direction on the sample stage 100. That is, during the process of the tool body 200 cutting the tissue block 10, the microscope objective 400 disposed opposite to the tool body 200 can avoid interfering with the movement of the tissue block 10. After the tissue section 11 is formed, the microscope objective 400 faces the cutting surface 211 of the tool body 200 to facilitate microscopic cutting of the tissue section 11 attached to the cutting surface 211.

[0033] Understandably, the microscope objective 400 can also be used for real-time microscopic observation of tissue sections 11, as well as for imaging and positioning of tissue sections 11 before microscopic cutting, to ensure the accuracy of microscopic cutting.

[0034] In some embodiments, such as Figure 3 As shown, the microscope objective 400 is also equipped with a liquid-drip device 410, which is used to drip liquid to cool the tissue section 11 and prevent heat accumulation on the blade tip 210 caused by long-term cutting.

[0035] Another embodiment of this application discloses a collection system for laser micro-cutting; please refer to [link to relevant documentation]. Figure 1 and Figure 2 It includes the aforementioned cutting tool for laser micro-cutting, support 800, gas guiding assembly 500, and cell sieve 600. The exhaust fan 300 and the cutting tool are both fixedly mounted on the support 800. The gas guiding assembly 500 is used to connect the exhaust end of the exhaust fan 300 with the first air outlet 230 of the cutting tool body 200, so that the exhaust fan 300 can draw gas from the first air inlet 220 and form a negative pressure area. Specifically, the gas guiding assembly 500 is provided with a gas guiding chamber 503. The gas guiding assembly 500 includes a first end 501 and a second end 502. The gas guiding chamber 503 connects the first end 501 and the second end 502. The first end 501 is connected to the first air outlet 230, and the second end 502 is connected to the exhaust fan 300. Thus, the gas guiding assembly 500 connects the airflow channels in the exhaust fan 300 and the cutting tool body 200.

[0036] Please see Figure 1 and Figure 5 The cell sieve 600 is disposed in the air guiding cavity 503 of the air guiding assembly 500. The cell sieve 600 includes sieve holes with a certain aperture size. The cell sieve 600 is disposed in the air guiding assembly 500 along a direction perpendicular to the gas flow. The edge of the cell sieve 600 is tightly attached to the inner wall of the air guiding cavity 503.

[0037] Specifically, after the microscope objective 400 performs micro-cutting on the tissue section 11 located on the blade 211, the cut target single cells or cell groups will enter the airflow channel of the blade body 200 under negative pressure, and then enter the air guide cavity 503. When flowing through the cell sieve 600, the cell sieve 600 intercepts those larger than its sieve aperture, thereby achieving the screening of target single cells or target cell groups.

[0038] Furthermore, the cell sieve 600 is provided with two or more layers, and the cell sieve 600 is arranged sequentially along the air flow direction in the air guide cavity 503 according to the sieve aperture size from large to small.

[0039] In other words, in a specific embodiment of this application, such as Figure 1 and 5 As shown, the diameter of the target single cell or target cell ring obtained by microdissection using the microscope objective 400 is approximately 20 μm. At this time, the cell sieve 600 has two layers with different pore sizes: one layer has a pore size of 100 μm, and the other layer has a pore size of 10 μm. The 100 μm pore size cell sieve 600 is positioned upstream of the 10 μm pore size cell sieve 600. Thus, the target single cell or target cell obtained by microdissection... The cells pass through a 100μm pore size cell sieve 600 and are intercepted and captured by a 10μm pore size cell sieve 600. Larger tissue fragments generated during microdissection are intercepted by the 100μm pore size cell sieve 600. Smaller cells or cell groups generated during microdissection pass through the 100μm pore size cell sieve 600 and the 100μm pore size cell sieve 600 in sequence and flow into the subsequent gas path, thereby achieving rapid and effective collection of target single cells or target cell groups.

[0040] It is worth noting that the blade used for laser micro-cutting, by setting an airflow channel inside the blade body 200, allows the tissue slice 11 to be stably attached to the blade surface 211 of the blade body 200 under controllable negative pressure. This significantly reduces slice displacement, warping, and detachment caused by cutting vibration, fluid disturbance, or laser scanning, and improves the spatial stability and positioning accuracy of the tissue slice 11 during the laser micro-cutting process. Simultaneously, the collection system for laser micro-cutting integrates the air guide component 500 between the airflow channel and the exhaust fan 300 with the cell collection path after laser micro-cutting, and sets a cell sieve 6 within the air guide cavity 503 of the air guide component 500. This invention enables single cells or cell populations cut by laser microscopy to be directionally transported along a predetermined path under negative pressure and retained with size selectivity. This avoids the problem of traditional electrostatic collection caps blocking the ultraviolet laser cutting light path due to their large size, and realizes long-distance, non-contact cell collection. The integrated design of this application improves the recovery rate and purity of target cells without introducing additional machinery to contact the target single cells and target cell populations, reduces the risk of sample cross-contamination, and enables the laser microscopy and cell collection process to be completed continuously and stably. This improves the applicability and reproducibility of the system in high-precision single cell acquisition and subsequent space omics and molecular analysis.

[0041] In some embodiments, such as Figure 1 , Figure 2 and Figure 6As shown, the collection system for laser micro-dissection also includes a cell sieve filter 700, which is fixedly mounted on the frame. The cell sieve filter 700 has a receiving element 710 at its lower part, and a receiving cavity 711 within the receiving element 710. The cell sieve filter 700 has a second air inlet 720 on its outer wall, which connects to the receiving cavity 711. The cell sieve filter 700 has a blocking element 730 at its upper part, which extends downwards into the receiving cavity 711. An annular space is formed between the blocking member 730 and the wall of the accommodating cavity 711; the blocking member 730 extends into the lower end of the accommodating cavity 711 and has a second air outlet 731, which penetrates the blocking member 730 and connects to the outer wall of the top of the cell sieve filter 700; the first air outlet 230 of the tool body 200 is connected to the second air inlet 720, and the exhaust fan 300 is connected to the second air outlet 731. Thus, the cell sieve filter 700 is connected in series in the air passage between the tool body 200 and the exhaust fan 300.

[0042] Accordingly, the air guiding assembly 500 includes a first air guiding pipe 510 and a second air guiding pipe 520; one end of the first air guiding pipe 510 serves as the first end 501 of the air guiding assembly 500 and is connected to the first air outlet 230, and the other end is connected to the second air inlet 720; one end of the second air guiding pipe 520 serves as the second end 502 of the air guiding assembly 500 and is connected to the exhaust fan 300, and the other end is connected to the second air outlet 731. Thus, the knife body 200, the cell sieve filter 700 and the exhaust fan 300 are connected in series, and the aforementioned cell sieve 600 is disposed inside the first air guiding pipe 510.

[0043] Understandably, when the microscope objective 400 microscopically cuts the tissue section 11 on the blade 211, cells or cell groups smaller than the pore size of all cell sieves 600 generated during the micro-cutting process are carried by the airflow through each cell sieve 600 and flow into the cell sieve filter 700 through the second air inlet 720. The annular space formed by the blocking member 730 and the cavity wall of the receiving cavity 711 guides the airflow to generate a high-speed rotating cyclone. Under the centrifugal force of the cyclone, these cells or cell groups are thrown towards the cavity wall of the receiving cavity 711, thus losing the kinetic energy previously pulled by the airflow, and can then fall along the cavity wall of the receiving cavity 711 and gather at the bottom of the receiving cavity 711. Thus, the airflow is initially purified by the cell sieve filter 700. Subsequently, the initially purified airflow flows out along the second air outlet 731 at the bottom of the blocking member 730.

[0044] Further, please see Figure 6The cell sieve filter 700 also includes a filter element 740, which has multiple micropores. The filter element 740 is disposed at the second air outlet 731. The micropores can block cells and cell groups. When the airflow that has been initially purified flows to the second air outlet 731, the micropores on the filter element 740 block the remaining cells and cell groups carried in the airflow within the receiving cavity 711, preventing them from flowing out from the second air outlet 731.

[0045] In some embodiments, see Figure 7 To prevent cells and other impurities deposited at the bottom of the accommodating cavity 711 from being swept up by the high-speed rotating cyclone above, the cell sieve filter 700 has a baffle 750 on the cavity wall of the accommodating cavity 711. The baffle 750 is located between the blocking member 730 and the bottom of the accommodating cavity 711. The edge of the baffle 750 is in close contact with the cavity wall of the accommodating cavity 711. The baffle 750 gradually extends downward from the edge to the center and forms a through hole at the center. Thus, the baffle 750 can guide the cells and cell groups thrown towards the cavity wall by the cyclone to the bottom of the accommodating cavity 711, and can also effectively isolate the high-speed rotating airflow above, preventing cells and other impurities that have settled at the bottom of the accommodating cavity 711 from being swept up again by the high-speed airflow above.

[0046] This application also discloses the workflow of a collection system for laser micro-cutting, the specific steps of which are as follows: The tissue block 10 is fixed on the sample stage 100; The exhaust fan 300 is activated to create a negative pressure area at the first air inlet 220 of the cutter body 200; The sample stage 100 is controlled to move the tissue block 10 so that the blade 210 can thinly slice the tissue block 10 and form a tissue slice 11. The tissue slice 11 is flatly attached to the blade 210 under the action of the negative pressure area. Before guiding the ultraviolet laser to perform micro-cutting of the tissue section 11, the microscope objective 400 is adjusted to image and position the tissue section 11. The ultraviolet laser is guided through the microscope objective 400 to the tissue slice 11 for microscopic cutting. The single cells or cell groups formed by the microscopic cutting of the tissue slice 11 enter the first suction tube under the action of the negative pressure area. The cell sieve 600 in the first suction tube screens the target single cells or target cell groups.

[0047] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of this application. Furthermore, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other.

Claims

1. A knife for laser microdissection, characterized in that include: A sample stage is movable, and a tissue block is fixed on one side of the sample stage in a first direction. The movement of the sample stage can drive the tissue block to move. The tool body is fixedly disposed in a first direction on the sample stage; the tool body includes a cutting head and an airflow channel, the airflow channel passes through the tool body and forms a first air inlet and a first air outlet on the surface of the tool body, the cutting head is used to cut tissue blocks to form tissue sections, the cutting head is disposed at the port of the first air inlet, the cutting head is located outside the first air inlet and is inclined relative to the sample stage; A blower is connected to the first air outlet of the cutter body to extract gas. A microscope objective is disposed on one side of the tool body. The microscope objective is used to conduct ultraviolet laser cutting light path to micro-cut tissue sections to form single cells or cell groups.

2. The cutting tool for laser micro-cutting according to claim 1, characterized in that, The blade head includes a blade surface located on one side of the blade head in a first direction. The sample stage is capable of moving the tissue block and bringing it into contact with the blade head. The blade head cuts the tissue block to form tissue sections, and the tissue sections are located on the blade surface.

3. The cutting tool for laser micro-cutting according to claim 2, characterized in that, The first air inlet is located in the first direction of the cutter head.

4. The cutting tool for laser micro-cutting according to claim 2, characterized in that, The microscope objective is positioned opposite to the tool body, with the lens of the microscope objective facing the cutting surface of the tool body. Both the microscope objective and the tool body are positioned in a first direction on the sample stage.

5. The cutting tool for laser micro-cutting according to claim 1, characterized in that, The microscope objective is used for real-time microscopic observation of tissue sections and for imaging and positioning of tissue sections before microscopic cutting.

6. A collection system for laser micro-cutting, characterized in that, include: The cutting tool for laser micro-cutting as described in any one of claims 1 to 5; A gas guiding assembly is provided with a gas guiding chamber. The gas guiding assembly includes a first end and a second end. The gas guiding chamber connects the first end and the second end. The first end is connected to the first air outlet of the tool body, and the second end is connected to the exhaust fan. The exhaust fan is used to extract gas sequentially through the airflow channels of the gas guiding assembly and the tool body. A cell sieve, which is disposed in the air-guiding assembly and separates the air-guiding chamber, is used to screen for target single cells or target cell populations.

7. The collection system for laser micro-cutting according to claim 6, characterized in that, The cell sieve has two or more layers, and the cell sieves are arranged sequentially along the airflow direction in the air guide cavity according to the sieve aperture size from large to small.

8. The collection system for laser micro-cutting according to claim 6, characterized in that, It also includes a cell sieve filter, which includes a receiving member with a receiving cavity inside; the cell sieve filter has a second air inlet on its outer wall, which is connected to the receiving cavity; the cell sieve filter also includes a blocking member that extends into the receiving cavity, and a second air outlet at one end of the blocking member that extends into the receiving cavity, which penetrates the blocking member and is connected to the outer wall of the cell sieve filter; the first air outlet of the blade body is connected to the second air inlet, and the exhaust fan is connected to the second air outlet.

9. The collection system for laser micro-cutting according to claim 8, characterized in that, The cell sieve filter further includes a filter element having multiple micropores, which is disposed at the second air outlet. The micropores are capable of blocking cells and cell clusters.

10. The collection system for laser micro-cutting according to claim 8, characterized in that, The air guiding assembly includes a first air guiding pipe and a second air guiding pipe; one end of the first air guiding pipe serves as the first end of the air guiding assembly and is connected to the first air outlet, and the other end is connected to the second air inlet; one end of the second air guiding pipe serves as the second end of the air guiding assembly and is connected to the exhaust fan, and the other end is connected to the second air outlet.

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