A cell block automatic preparation machine

Abstract

The present invention relates to an automatic cell block preparation machine and method. The automatic cell block preparation machine comprises a main body and a centrifugal component, a temperature control component, a mechanical arm, a liquid circuit system and a desktop host arranged on the main body. Based on the cell block preparation machine platform, cells in a cytological specimen are separated and enriched on a plane through the centrifugal action of a cell block preparation tube and a matrix, and are automatically prepared into cell blocks, thereby realizing the automation, standardization and high quality of cell block preparation.

Description

Technical Field

[0001] The present invention relates to the technical field of medical equipment, and in particular to an automatic cell block preparation machine and method. Background Art

[0002] Cytopathology involves the analysis of cells or small groups of cells to diagnose a variety of diseases and assess health.

[0003] The most commonly used technique in cytopathology is cell smear. Cell smear is to spread cells or small cell clusters on a glass slide, stain them with diagnostic reagents, and observe the cell morphology under a microscope to diagnose a variety of diseases or assess health conditions. The cell smear method has obvious limitations, mainly manifested in: cell smears cannot be replicated. Once a cell smear is stained with a diagnostic reagent, it is difficult to perform other staining and testing at the same time; it is difficult to perform immunohistochemistry and molecular pathology tests; factors such as blood, protein precipitation, mucus interference, and cell accumulation make the cell morphology and structure unclear; cells are twisted and deformed during the preparation process and are difficult to interpret; they cannot be stored for a long time, etc.

[0004] Cell blocks are the bridge from cytopathology to precision medicine. After dehydration and paraffin embedding, cell blocks are made into cell wax blocks, which are cut into cell sections. After further staining, cells are tested and diagnosed for diseases. Compared with cell smears, cell sections have many advantages: cells do not deform, the structure is clear, and they are easy to observe and interpret; the arrangement of cells can be clearly observed like biopsy tissue sections, so they are also called microbiopsies; cell blocks can cut out multiple sections of cells or small cell clusters for analysis; they are easy to perform special staining, immunocytochemical staining, and molecular pathology testing; and they can be stored for a long time for further diagnosis and research.

[0005] There are many methods for making cell blocks, which can be summarized as direct method and indirect method. The direct method is to use ordinary centrifuge tubes to centrifuge and then aspirate the supernatant, then remove the sediment, and then use conventional pathological techniques to make wax blocks; the indirect method is to add substances that can cause cells to aggregate into clusters to the cell sediment, so that the cells can be aggregated and then taken out to prepare cell blocks. However, these existing methods have different defects, among which the most obvious defects are: (1) all cell blocks are prepared manually one by one, which is time-consuming and labor-intensive; (2) the preparation of cell blocks lacks standardization, regularization and quality control; (3) it is not suitable for preparing cell blocks when the number of cells or cell clusters is small; (4) the position of the target cells (pathological cells) in the cell wax block is scattered and uncertain, making it difficult to cut the diseased cells when slicing; (5) the distribution area of ​​cells on the slice is small and the density is low, which makes diagnosis difficult; (6) the size and shape of the wax block are irregular. Summary of the invention

[0006] The present invention provides an automatic cell block preparation machine and method. The automatic cell block preparation machine is mainly composed of a body and a centrifugal assembly, a temperature control assembly, a liquid circuit assembly, a mechanical arm and a desktop host arranged on the body. The centrifugal assembly is located at the lower part of the body, the mechanical arm is located at the upper part, and the host is located at the front lower part. The centrifugal assembly includes a driving motor, a rotor, a hanging cup, an adapter and a cell block preparation tube. The rotor is provided with an embedding box clamp. The cell block preparation tube is a cylindrical hollow tube with two ends open, a large inner diameter at the upper end, a small inner diameter at the lower end or the same inner diameter at the upper and lower ends. The outer layer at both ends has an external thread, and the bottom cover of the preparation tube is flat-bottomed, and the inner side wall surface layer has an internal thread, which can be tightened and engaged with the external thread of the preparation tube to form a cylindrical closed space. The upper end of the cell block preparation tube is provided with a clamping shaft, and the bottom cover is provided with a clamping tooth. The bottom of the adapter is provided with a heating plate and a cell block preparation tube installation slot, which can realize heating / cooling of the cell block preparation tube, and support and fix the cell block preparation tube, and support the mechanical arm to remove the centrifuge tube cover when rotating the centrifuge tube. The temperature control component mainly has a bidirectional function of heating / cooling. The robotic arm is provided with a bayonet for clamping the cell block preparation tube, and is equipped with a piston, which can rotate and unscrew the cell block preparation tube and push out the cell block. The method of making cell blocks mainly uses a substance whose physical state (liquid and solid) changes with temperature as a matrix material, a liquid matrix as a medium for centrifugal separation of cells, and a solid matrix after cooling and solidification as a cell support embedding material. The cell support after centrifugal separation and enrichment is fixed in a specific space and prepared into a cell block. The melting point of the matrix is ​​in the range of 60℃~100℃, and the solidification point is in the range of 20℃~60℃. Applicable matrices include agar, agarose, gelatin and matrix glue. Through the centrifugal separation of the cell block preparation tube and the matrix, the cells are separated and enriched on a plane, and the diseased cells are distributed in the bottom layer.

[0007] The beneficial effects of the present invention are: (1) Through the automatic cell block preparation machine and the cell block preparation method, cytological specimens and tiny tissue fragments obtained by surgical methods can be automatically prepared into cell blocks.

[0008] (2) To achieve automation, standardization and quality control of cell block preparation in clinical practice.

[0009] (3) Using cell block preparation tubes and matrix materials, through the principle of centrifugation, a small amount of cells and tissue debris can be efficiently enriched on one plane of the matrix, thereby preparing trace cells into cell blocks.

[0010] (4) Through the centrifugal separation of the matrix material, the diseased cells and cell clusters (large cells, cells with a high nuclear-to-cytoplasmic ratio) are distributed in the bottom layer and are cut off first to make cell slices when sections are made, thereby increasing the diagnostic positivity rate.

[0011] (5) The slices made using this method have a large observable cell distribution area and high cell abundance, which improves the quality of diagnosis.

[0012] (6) The cell blocks prepared by this method have regular shapes and definite cell distribution positions. The cell sections prepared by this method are of high quality and easy to diagnose. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 It is a structural schematic diagram of an automatic cell block preparation machine of the present invention.

[0014] Figure 2 It is a schematic diagram of the structure of the centrifugal assembly of the present invention.

[0015] Figure 3 It is a schematic diagram of the structure of the cell block preparation tube of the present invention.

[0016] Figure 4 It is a schematic diagram of the adapter structure of the present invention.

[0017] Figure 5 It is a connection schematic diagram of the fluid path assembly of the present invention.

[0018] Figure 6 The figure is a flow chart of the method for preparing cell blocks of the present invention. DETAILED DESCRIPTION

[0019] In order to make the objectives, technical solutions and advantages of the present invention more clear, it should be noted that the directions or positional relationships indicated by terms such as "upper", "lower", "left", "right", "inside", and "outside" are based on the directions or positional relationships shown in the accompanying drawings, and are only for the convenience of description and simplified description, and do not refer to or imply that the device or component referred to must have a specific direction. In addition, in the description of the present invention, the terms "installation" and "connection" should be understood in a broad sense.

[0020] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments of the specification.

[0021] like Figure 1 As shown, an automatic cell block preparation machine of the present invention includes a main body 1 and main components such as a centrifugal assembly 2, a mechanical arm (including a moving mechanical arm 5, a liquid discharge mechanical arm 6 and a sample loading mechanical arm 7), a temperature control assembly 41 and a host 10 arranged in the main body, and also includes a reagent bottle 8, a sample loading needle cleaning device 9 and a storage tank 11. The main body 1 is a box structure, the centrifugal assembly 2 is located at the lower part of the main body, the mechanical arm is located at the upper part of the main body, and the desktop host 10 is located at the front lower part.

[0022] like Figure 2 As shown, the centrifuge assembly includes a driving motor 21, a rotor 22, a hanging cup 23, a cell block preparation tube 3 and an adapter 4; the rotor 22 is a disc-shaped structure, and an embedding box holder 24 is provided above it.

[0023] like Figure 3 As shown, the cell block preparation tube 3 is a cylindrical hollow tube with two ends open, with a large inner diameter at the upper end and a small inner diameter at the lower end or the inner diameters of the upper and lower ends are the same, and the outer surface layers at both ends have external threads, and the inner side wall surface layers of the bottom cover 31 and the top cover 32 of the cell block preparation tube 3 have internal threads, which can be tightened and engaged with the external threads of the cell block preparation tube 3 to form a cylindrical space. The bottom cover 31 is flat-bottomed, which is conducive to enriching cells on a plane after centrifugation. The upper end of the cell block preparation tube 3 is provided with a latch 34, and the bottom cover is provided with a latch tooth 33.

[0024] like Figure 4 As shown, a heating plate 41 and a cell block preparation tube 3 mounting slot 42 are provided at the bottom of the adapter 4 for supporting and fixing the cell block preparation tube 3 , which is conducive to moving the mechanical arm 5 to rotate and unscrew the bottom cover 31 of the cell block preparation tube 3 .

[0025] like Figure 5 As shown, the liquid circuit assembly includes a sample addition liquid circuit and a liquid discharge liquid circuit. The sample addition liquid circuit starts from the red blood cell lysis solution 81, the cell fixative solution 82 and the matrix 83 respectively, passes through the diaphragm liquid pump 84 and the solenoid valve 85, and is connected to the sample addition needle to drip the detection reagent into the cell block preparation tube 3; the liquid discharge liquid circuit starts from the cell block preparation tube 3, passes through the diaphragm liquid pump and the solenoid valve, and discharges the liquid in the cell block preparation tube 3 into the waste liquid bottle 86.

[0026] like Figure 6 As shown, the cell block preparation flow chart includes three specific implementation schemes ( Figure 6 a, 6b, 6c), Specific implementation plan 1 ( Figure 6 a): specimen collection S1, cell block preparation tube installation S2, centrifugation S3, removal of supernatant S4, red blood cell lysis S5, centrifugation S6, removal of supernatant S7, cell fixation S8, addition of matrix S9, centrifugation S10, matrix coagulation S11, removal of cell block preparation tube and pushing out cell block S12; Specific implementation plan II ( Figure 6 b): specimen collection S1, cell block preparation tube installation S2, centrifugation S3, removal of supernatant S4, red blood cell lysis S5, centrifugation S6, removal of supernatant S7, cell fixation S8, matrix liquefaction S9, centrifugation S10, matrix coagulation S11, removal of cell block preparation tube and pushing out cell block S12; Specific implementation plan three ( Figure 6 c): Specimen collection S1, cell block preparation tube installation S2, centrifugation S3, supernatant removal S4, red blood cell lysis S5, centrifugation S6, supernatant removal S7, cell fixation S8, matrix addition S9, centrifugation S10, cell block preparation tube removal S11, matrix solidification S12, sample collection S13.

[0027] The technical solution of the present invention is further illustrated by describing in detail the working process of the automatic cell block preparation machine.

[0028] Specific implementation plan 1 Figure 6 a).

[0029] (1) Specimen collection: Unscrew the top cap 32 of the cell block preparation tube 3 and add the cytology specimen into the cell block preparation tube 3.

[0030] (2) Installation of the cell block preparation tube 3 : The cell block preparation tube 3 containing the cytological specimen is installed in the adapter 4 , and the latching teeth 33 on the bottom cover 31 are inserted into the latching grooves 43 of the adapter 4 .

[0031] (3) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells at the bottom of the cell mass preparation tube 3.

[0032] (4) Removing the supernatant: The driving motor (21) rotates the cell block preparation tube (3) to the work station, and the liquid discharge robot arm 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant is sucked out by the diaphragm liquid pump.

[0033] (5) Red blood cell lysis (if necessary): Add red blood cell lysis buffer to the cell pellet, pipette to mix, and let stand for 5 minutes to lyse the red blood cells.

[0034] (6) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells at the bottom of the cell mass preparation tube 3.

[0035] (7) Removing the supernatant: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the liquid discharge robot 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant containing red blood cell fragments is sucked out by the diaphragm liquid pump.

[0036] (8) Cell fixation: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 moves to add the cell fixative 81 into the cell block preparation tube 3, and mixes by pipetting and blowing, and fixes the cells for 10 minutes.

[0037] (9) Adding matrix liquid: The driving motor rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 moves to add the pre-heated liquefied matrix into the cell block preparation tube 3 and mix it with the cells.

[0038] (10) Centrifugation: The cell block preparation tube 3 is centrifuged at 1500 RPM (400 g) for 5 minutes. Through the separation effect of the matrix, the cells are separated and enriched on a plane at the lower end of the matrix. The diseased cells are distributed in the lower layer due to their large size.

[0039] (11) Matrix solidification: The cell block preparation tube 3 is cooled by a refrigeration component to solidify the matrix, thereby fixing the separated cell supports at specific spatial positions of the matrix.

[0040] (12) Removing the cell block: The driving motor 21 rotates the cell block preparation tube to the work station, moves the robot arm 5 and docks its bayonet 51 with the clamping shaft at the upper end of the cell block preparation tube 3, applies a rotational force, rotates and removes the cell block preparation tube 3 in the adapter 4, and moves it to the top of the embedding box clamping position 24, and uses the piston 52 to push the cell block into the embedding box.

[0041] (13) The entire process is precisely controlled by the host computer 7.

[0042] Specific implementation plan 2 Figure 6 b): Scheme in which the cell block preparation tube 3 is pre-filled with matrix.

[0043] (1) Specimen collection: Unscrew the top cap 32 of the cell block preparation tube 3 and add the cytology specimen into the cell block preparation tube 3.

[0044] (2) Installation of the cell block preparation tube 3: Install the cell block preparation tube 3 containing the cytological specimen into the adapter 4 and insert the latch teeth 33 on the bottom cover into the latch groove 42 of the adapter.

[0045] (3) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells onto the top surface of the solid matrix in the cell block preparation tube 3.

[0046] (4) Removing the supernatant: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the liquid discharge robot 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant is sucked out by the liquid pump.

[0047] (5) Red blood cell lysis (if necessary): The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 adds red blood cell lysis solution to the cell pellet, blows and mixes it, and leaves it to stand for 5 minutes to lyse the red blood cells.

[0048] (6) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells on the top surface of the solid matrix in the cell block preparation tube 3.

[0049] (7) Removing the supernatant: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the liquid discharge robot 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant containing red blood cell fragments is sucked out by the liquid pump.

[0050] (8) Cell fixation: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 5 moves to add the cell fixative 81 into the cell block preparation tube 3, and mixes by pipetting, and fixes the cells for 10 minutes.

[0051] (9) Matrix liquefaction: The matrix in the cell block preparation tube 3 is heated to 75 degrees by the heating plate 41 at the bottom of the adapter 4 to liquefy the matrix.

[0052] (10) Centrifugation: Cell block preparation tube 3 is centrifuged at 1500 RPM (400 g) for 5 minutes. The cells are separated and enriched on a plane at the lower end of the matrix through the separation effect of the matrix. The diseased cells are distributed in the lower layer due to their large size.

[0053] (11) Matrix solidification: The cell block preparation tube 3 is cooled by the cooling effect of a centrifuge to solidify the matrix, thereby fixing the separated cells to a specific spatial position of the matrix.

[0054] (12) Removing the cell block: The driving motor 21 rotates the cell block preparation tube 3 to the work station, moves the robot arm 5 and docks its bayonet 51 with the clamping shaft 34 at the upper end of the cell block preparation tube 3, applies a rotational force, rotates and removes the cell block preparation tube 3 in the adapter 4, and moves it to the top of the embedding box clamping position 24, and uses the piston 52 to push the cell block into the embedding box.

[0055] (13) The entire process is precisely controlled by the host computer 7.

[0056] Specific implementation plan three ( Figure 6 c): In order to make the device easy to manufacture, the step of removing the cell block is done manually.

[0057] (1) Specimen collection: Unscrew the top cap 32 of the cell block preparation tube 3 and add the cytology specimen into the cell block preparation tube 3.

[0058] (2) Installation of the cell block preparation tube 3: The cell block preparation tube 3 containing the cytological specimen is installed in the adapter 4.

[0059] (3) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells onto the bottom cover of cell block preparation tube 3.

[0060] (4) Removing the supernatant: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the liquid discharge robot 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant is sucked out by the liquid pump.

[0061] (5) Red blood cell lysis (if necessary): The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 adds 2 ml of red blood cell lysis solution to the cell pellet, blows and mixes, and leaves it to stand for 5 minutes.

[0062] (6) Centrifugation: Centrifuge at 1500 RPM (400 g) for 5 minutes to separate and enrich the cells at the bottom of the cell mass preparation tube 3.

[0063] (7) Removing the supernatant: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the liquid discharge robot 6 moves to insert the sample injection needle into the supernatant of the cell block preparation tube 3, and the supernatant containing red blood cell fragments is sucked out by the liquid pump.

[0064] (8) Cell fixation: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 moves to add the cell fixative 81 into the cell block preparation tube 3, and mixes by pipetting and blowing, and fixes the cells for 10 minutes.

[0065] (9) Adding matrix liquid: The driving motor 21 rotates the cell block preparation tube 3 to the work station, and the sample loading robot 7 moves to add the pre-heated liquefied matrix into the cell block preparation tube and mix it with the cells.

[0066] (10) Centrifugation: Cell block preparation tube 3 is centrifuged at 1500 RPM (400 g) for 5 minutes. The cells are separated and enriched on a plane at the lower end of the matrix through the separation effect of the matrix. The diseased cells are distributed in the lower layer due to their large size.

[0067] (11) Take out the cell block preparation tube: Manually take out the cell block preparation tube and place it on the test tube rack.

[0068] (12) Matrix solidification: Place the cells at room temperature for 20 minutes or in a refrigerator at 4 degrees for 10 minutes to allow the matrix to solidify and fix the separated cells in a specific spatial position.

[0069] (13) Sampling: Unscrew the bottom cap at the lower end of the cell block preparation tube, push out the cell end of the solid matrix with a cotton swab, and place the cut cell block into the embedding box.

[0070] It should be noted that the above description of the specific embodiments of the present invention is only for illustrating the technical route and features of the present invention, and its purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly, but the present invention is not limited to the above specific implementation methods. Any changes or modifications made within the scope of the claims of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A cell block automatic preparation machine, characterized in that: It includes an automatic cell block preparation machine and a method for preparing cell blocks based on the preparation machine; The automatic cell block preparation machine is characterized by comprising a body and a centrifugal assembly, a temperature control assembly, a mechanical arm, a fluid path system and a desktop host arranged on the body, wherein the centrifugal assembly is located at the lower part of the body, the temperature control assembly is located at the bottom, the mechanical arm is located at the upper part, and the host is located at the lower front part; The centrifugal assembly is characterized in that it includes a driving motor, a rotor, a hanging cup, an adapter and a cell block preparation tube; the upper end of the cell block preparation tube is provided with a clamping position, the bottom cover is provided with a latching tooth, the bottom of the adapter is provided with a heating plate and a cell block preparation tube installation slot, which cooperates with the latching tooth on the bottom cover to support and fix the cell block preparation tube, and is used for the robot arm to rotate and remove the cell block preparation tube; the cell block preparation tube is installed in the adapter, and then hung on the rotor through the hanging cup, and the driving motor drives the rotor to rotate; The temperature control component is characterized by having two functions: heating and cooling; The robotic arm is characterized in that it comprises a sample loading robotic arm, a liquid discharge robotic arm and a moving robotic arm, wherein the sample loading robotic arm adds detection reagent to the cell preparation tube through a sample loading needle, the liquid discharge robotic arm discharges the supernatant of the cell preparation tube through the sample loading needle, and the moving robotic arm clamps and moves the cell preparation tube, rotates and unscrews the bottom cover of the cell preparation tube and pushes out the cell block; The method is characterized in that, relying on the automatic cell block preparation machine and the matching cell block preparation tube and matrix material, the cell components in the cytological specimen are separated and enriched, and the detection reagent is automatically added to prepare the cell block.

2. The automatic cell block preparation machine according to claim 1, characterized in that: The centrifugal assembly has a rotor positioning function, which can rotate any cell preparation tube to the target position after the centrifugation is completed.

3. The automatic cell block preparation machine according to claim 1, characterized in that: Methods for implementing the rotor positioning function include: (1) using a drive motor with a positioning function; and (2) installing a rotor positioning device on the drive motor.

4. The automatic cell block preparation machine according to claim 1, characterized in that The rotor is a disc-shaped structure with an embedding cassette holder on top.

5. The automatic cell block preparation machine according to claim 1, characterized in that: The cell block preparation tube is a cylindrical hollow tube with openings at both ends. The bottom cover is flat and can be screwed and engaged with the lower end of the cell block preparation tube to prevent liquid leakage. The shape includes: (1) the inner diameter of the upper end is large and the inner diameter of the lower end is small; (2) the inner diameters of the upper and lower ends are the same.

6. The automatic cell block preparation machine according to claim 1, characterized in that: The matrix material is a material that solidifies into a solid state at room temperature and melts into a liquid state after heating. Its melting point is in the range of 60°C to 100°C and its solidification point is in the range of 20°C to 60°C.

7. The automatic cell block preparation machine according to claim 1, characterized in that: The matrix is ​​liquefied by heating, the liquid matrix serves as a medium for centrifugal separation of cells, and the solid matrix after cooling serves as a supporting embedding material for the cells. The cells enriched by centrifugal separation are prepared into cell blocks. The specific method includes: (1) mixing the liquid matrix with the cells and then centrifuging to prepare cell blocks; (2) first centrifuging the cells to the bottom of a cell preparation tube, and then adding the matrix to the cell sediment to prepare cell blocks; (3) first centrifuging the cells to the surface of the solid matrix, and then heating the liquefied matrix and centrifuging to prepare cell blocks.

8. The automatic cell block preparation machine according to claim 1, characterized in that: Through the centrifugal separation of the cell block preparation tube and the matrix, the cells are enriched on one plane, and the cell clusters and large cells are distributed in the lower layer.

Images