CN113390699B - Device and method for preparing cell wax block in stamping mode - Google Patents

Abstract

The invention relates to the technical field of medical electronic instruments, in particular to a device and a method for preparing cell wax blocks in a stamping mode. The device and the method for preparing the cell wax block in the stamping mode have the advantages of convenient, stable and smooth preparation process, strong applicability, no influence by the size and the number of cells of cell sap to be prepared, low technical requirements on operators, low heating energy consumption, capability of ensuring full reaction by repeatedly stamping continuously added new reagents, accelerated reaction time and shortened preparation time.

Description

A punching type device and method for preparing cell wax blocks

technical field

The invention relates to the technical field of medical electronic equipment, in particular to a punching type device and method for preparing cell wax blocks.

Background technique

The cell wax block is to extract the cells in the liquid and then fix them with paraffin. The experience level of the operator is directly related, otherwise it is likely that the quality of the wax blocks will be uneven. In addition, because formalin, xylene and other reagents need to be added during the operation, a little carelessness will bring harm to the health of the operator. to hurt.

In prior art such as patent "CN2014101669573 device and method for automatically preparing cell wax block" and "CN2014101643499 pressurized reaction device for preparing cell wax block", a method and device for automatically preparing cell wax block are proposed. A series of steps such as fixation, dehydration, transparency, staining, wax dipping, and cooling are all automatically completed in the instrument. After the operation is completed, a wax-soaked cell block can be formed in the test tube. After the cell block is taken out, it is placed in an embedding frame for packaging. After burying, the cells can be sent for sectioning after the wax block. This method and device solve the shortcomings of manual operation.

But find that this device and method also have some problems and shortcoming in practice. The problem is especially pronounced when the body fluid contains many cells. After the body fluid is poured into a test tube with a semi-permeable membrane, the reagents, water and small molecules in it will pass through the semi-permeable membrane, while the cells will stay on the semi-permeable membrane. When a considerable amount of cells accumulate on the semi-permeable membrane, it will hinder the filtration of liquid and small molecules and make the subsequent process slower and slower. At this time, it is necessary to speed up the process by applying negative pressure. If the negative pressure applied is too small, the effect will not be obvious; if it is too large, it will cause slightly larger cells to embed in the holes in the semi-permeable membrane, although the cells that partially block the semi-permeable membrane can be repressed by reversely applying positive pressure in the test tube. In the cell fluid, there will still be a small amount of cells that block the semi-permeable membrane and remain on the semi-permeable membrane. After repeated operations like this for many times, the filtration characteristics of the semi-permeable membrane will eventually disappear, resulting in poor final results. In addition, due to the wax immersion process, the paraffin wax will solidify if the temperature is lower than 40°C, so the test tube, pressurization and extraction device need to maintain a certain temperature during the wax immersion process. The test tube is disposable, so it's not a big problem. However, due to the cyclic operation of the pressurization and extraction devices, the paraffin attached to the inner wall will accumulate more and more after working for a long time. In order to ensure smooth work, the pressurization and extraction devices need to be heated during the rest of the fixation, dehydration, transparency, dyeing and other processes, resulting in inconvenience and excessive energy consumption.

For the above technical problems, it needs to be improved.

Contents of the invention

The object of the present invention is to provide a punching-type device and method for preparing cell wax blocks to address the defects of the prior art.

In order to achieve the above object, the present invention adopts the following technical solutions:

A stamping type device for preparing cell wax blocks, including a turntable, a test tube, a first stamping device, a second stamping device, a liquid suction device, and a sleeve heating device;

The turntable has a plurality of symmetrically arranged fixing holes for fixing the test tubes;

The inner bottom of the test tube is inlaid with a metal sheet, and the surface of the metal sheet is smooth. The bottom part of the test tube where the metal sheet is located is a detachable structure or can be cut and separated. The test tube wall has a symmetrically arranged first stamping Pipeline, the second stamping pipeline, the outlet end of the stamping pipeline is set on the test tube wall above the metal sheet inside the test tube, the entrance end of the stamping pipeline is set on the outer side of the test tube wall or the top of the test tube, and the stamping tube There is a quick-plug interface at the entrance of the road, and a one-way valve is arranged in the stamping pipeline;

The first stamping device is used to connect the first stamping pipeline and stamp reagents into the first stamping pipeline, including several reagent bottles, a first conduit, a first pressurizing pump, a solenoid valve, and a first mechanical arm. The first pressurizing pump is installed on the first conduit pipeline, and the first conduit communicates with the reagents in several reagent bottles through the solenoid valve, and the first mechanical arm controls the first conduit to connect with the quick-plug interface of the first punching pipeline;

The second stamping device is used to connect the second stamping pipeline and stamp liquid paraffin into the second stamping pipeline, including a metal paraffin pool, a second conduit, a second booster pump, and a second mechanical arm. The booster pump is installed on the second conduit pipeline, one end of the second conduit is connected to the metal paraffin pool, and the second mechanical arm controls the second conduit to connect with the quick-plug interface of the second stamping pipeline;

The liquid suction device is used to absorb liquid from the test tube, including a collection bottle, a suction pipe, a working pump, and a third mechanical arm. The working pump is installed on the pipette pipeline, and the suction pipe communicates with the collection bottle. The third mechanical arm controls the pipette to extend into or out of the test tube;

The sleeve heating device is arranged under the turntable for heating the test tube, including a liftable heating sleeve, the inner shape of the heating sleeve container is adapted to the outer shape of the test tube to wrap the bottom of the test tube, and the heating sleeve Has a heating film inside.

As a preferred solution, a metal mesh is attached to the outside of the test tube.

As a preferred solution, the bottom of the test tube is a circular truncated structure, and the test tube shrinks from the mouth of the test tube to the circular truncated bottom.

As a preferred solution, a positioning block is provided at the middle and lower part of the outer side of the test tube, and a positioning protrusion matching the positioning block of the test tube is provided at the top of the heating sleeve.

As a preferred solution, the device for automatically preparing cell wax blocks also has a liquid level detector for detecting the liquid level in the test tube.

As a preferred solution, the device for automatically preparing cell wax blocks also has a liquid flow rate sensor for sensing the liquid flow rates in the first conduit and the second conduit.

A method for punching a device for preparing cell wax blocks, comprising the steps of:

S1. Take a sample, put the cell solution of the cell wax block to be prepared into a test tube, and place the test tube in the fixing hole of the turntable to fix;

S2. Form cell clumps, gather dense substances at the bottom of the test tube by centrifugation to form cell clumps;

Among them, centrifugation means that the turntable rotates at 1000-2000 rpm for 1-2 minutes, and then according to the process, the suction device sucks out the supernatant liquid or reagents and liquid paraffin in the test tube;

S3. Fixation, the solenoid valve is connected to the reagent for fixation, and the process of stamping the reagent, natural precipitation, repeated stamping and precipitation, and centrifugation is repeated. The process of fixation lasts 1.5-2 hours. This process ensures that the reagent in the test tube is higher than the cells at the bottom lump 5cm;

Among them, the reagent for fixation refers to neutral formalin with a concentration of 10%; the stamping reagent refers to the first stamping device docking the first stamping pipeline and stamping the reagent into the first stamping pipeline to drive the liquid in the test tube to flow and disperse it. Cell clumps; natural precipitation refers to the natural precipitation of the cell liquid in the test tube for 5-15 minutes; repeated stamping precipitation refers to the process of stamping reagent/liquid paraffin and natural precipitation repeated 2-4 times;

S4. Dehydration, including primary dehydration and secondary dehydration. For the primary dehydration, the solenoid valve communicates with the reagent for primary dehydration, and repeats the process of stamping reagents, natural precipitation, repeated stamping precipitation, and centrifugation. The primary dehydration process lasts 1- 1.5 hours; for the second dehydration, the solenoid valve is connected to the reagent for the second dehydration, and repeats the process of stamping reagents, natural precipitation, repeated stamping precipitation, and centrifugation. The secondary dehydration process lasts 1-1.5 hours;

Among them, the reagent for primary dehydration refers to alcohol with a concentration of 95%, and the reagent for secondary dehydration refers to absolute alcohol;

S5. Staining, the solenoid valve is connected to the reagent for dyeing, and the process of punching, natural precipitation, and repeated punching and precipitation is carried out. The dyeing process lasts 15-20 minutes. This process ensures that the reagent in the test tube is 2cm higher than the cell mass at the bottom;

Among them, the staining reagent refers to eosin;

S6. Transparent, the electromagnetic valve is connected to the transparent reagent, and the process of stamping reagent, natural precipitation, repeated stamping precipitation, and centrifugation is repeated. The transparent process lasts 25-30 minutes. This process ensures that the reagent in the test tube is higher than the cells at the bottom. lump 5cm;

Among them, the reagent for transparency refers to xylene;

S7. Soak in paraffin, repeat the process of punching liquid paraffin, heating the test tube, natural precipitation, repeated stamping and precipitation, removing the test tube from heating, and centrifuging. The process lasts for 1-1.5 hours, and the temperature of the test tube is 60-70°C , this process ensures that the liquid paraffin in the test tube is 5cm higher than the cell mass at the bottom;

Among them, stamping liquid paraffin means that the second stamping device is connected to the second stamping pipeline and stamps liquid paraffin into the second stamping pipeline; test tube heating means that while stamping liquid paraffin, the sleeve heating device controls the heating sleeve to rise to the heating sleeve. The tube wraps the bottom of the test tube, and the heating film in the heating sleeve starts to heat; before the test tube is separated from the heating finger, the sleeve heating device controls the heating sleeve to drop until it is separated from the test tube;

S8. Take out the test tube, disassemble or cut and separate the bottom of the test tube wall, and peel off the metal sheet to obtain the cell wax block.

As a preferred solution, the process of repeated stamping and precipitation also includes the step of: before stamping the reagent/liquid paraffin, detect the liquid level in the test tube, if the liquid level is less than 2-3cm away from the test tube mouth, the liquid suction device will remove the liquid in the test tube. The reagent/liquid paraffin is sucked out, and the liquid height in the control test tube is 4-5cm lower than the test tube mouth.

As a preferred solution, when the reagent or liquid paraffin is punched, the corresponding flow velocity in the first conduit or the second conduit is 0.2-0.3 m/s.

Compared with the prior art, the present invention has a stamping-type device and method for preparing cell wax block, the preparation process is convenient, stable, smooth, and has strong applicability, and is not affected by the size and quantity of the cells to be prepared, and has great impact on The technical requirements of the operator are low, and the heating energy consumption is low. Repeated punching and continuous addition of new reagents can ensure sufficient reaction, while speeding up the reaction time and reducing the preparation time.

Description of drawings

FIG. 1 is a schematic structural view of a stamping-type device for automatically preparing cell wax blocks according to Embodiment 1 of the present invention;

2 is a schematic perspective view of the structure of the test tube of the stamping type automatic preparation of cell wax block according to Embodiment 1 of the present invention;

3 is a schematic front view of the structure of the test tube of the stamping type automatic preparation of cell wax block according to Embodiment 1 of the present invention;

4 is a schematic cross-sectional view of the structure of the test tube of the stamping type automatic preparation of cell wax block according to the first embodiment of the present invention;

5 is a schematic structural view of the heating sleeve of the stamping type automatic preparation of cell wax block according to Embodiment 1 of the present invention;

Fig. 6 is a schematic flow chart of a stamping-type device for automatically preparing cell wax blocks according to Embodiment 1 of the present invention;

Among them: 1. Turntable; 11. Fixing hole; 2. Test tube; 21. The first stamping pipeline; 22. The second stamping pipeline; 23. Quick plug interface; 24. Metal sheet; 25. Positioning block; 1. Punching device; 31. Reagent bottle; 4. Second punching device; 41. Metal paraffin pool; 5. Sleeve heating device; 51. Heating sleeve; 52. Positioning protrusion; 53. Heating film; Device; 61. Collection bottle.

detailed description

Embodiments of the present invention are described below through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation modes, and various modifications or changes can be made to the details in this specification based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that, in the case of no conflict, the following embodiments and features in the embodiments can be combined with each other.

Embodiment one:

As shown in Figures 1-6, this embodiment proposes a device and method for automatically preparing cell wax blocks by stamping, so that the automatic operation of cell mass fixation, dehydration, transparency, dyeing, wax dipping, cooling, etc. Complete in the designed test tube 2 without the need for a semi-permeable membrane to cooperate with positive and negative pressure.

Specifically, the device for automatically preparing cell wax blocks includes a turntable 1 , a test tube 2 , a first punching device 3 , a second punching device 4 , a liquid suction device 6 , and a sleeve heating device 5 .

Among them, the turntable 1 has a plurality of symmetrically arranged fixing holes 11 for fixing the test tubes 2. The turntable 1 can be opened with 4, or 8, 16 and other fixing holes 11 according to the size and scale, that is, it can be placed at one time. The corresponding number of test tubes 2 should be placed symmetrically to prevent deviation during centrifugation.

The inner bottom of the test tube 2 is inlaid with a metal sheet 24. The shape of the metal sheet 24 fits the bottom of the test tube 2. The surface of the metal sheet 24 is required to be smooth so that the centrifuged cell mass can be attached to its surface. After entering the stamping process at the same time, The cell clumps will be broken up and will not always adhere to the metal sheet 24; the bottom of the test tube 2 where the metal sheet 24 is located is a detachable structure or can be cut and separated, and the detachable structure can be threaded or interference fit. Inserting, clamping, etc., can be cut and separated, and the material of the test tube 2 is made of flexible materials, such as plastic, which is consistent with the test tube 2 used in ordinary experiments. It can be cut for one-time use, and can be disassembled and reused according to needs. When the wax dipping step is completed 1. After the paraffin in the test tube 2 is cooled, the operator can conveniently disassemble the test tube 2, peel off the metal sheet 24, and obtain the prepared cell wax block. The metal sheet 24 can be designed to be embedded directly as an embedding frame, or transferred to the embedding frame as a transfer for the next slicing operation. At the same time, when the metal sheet 24 is heated during the preparation process, it can improve the local heat conduction at the bottom and stabilize the cell mass The local preparation temperature requirements of the block are improved, and the heat utilization efficiency is improved. The test tube 2 has a positioning block 25 in the middle and lower part of the outer side, which is used for positioning with the sleeve heating device 5 . The fixing hole 11 of the turntable 1 is fitted with an avoidance hole for the positioning block 25 to pass through. Further, the bottom of the test tube 2 is designed as a circular table structure, and the test tube 2 is narrowed from the mouth of the test tube 2 to the bottom circular table. The diameter of the mouth of the test tube 2 is large, and then it is continuously inclined downwards, and the diameter is slightly reduced, so as to facilitate the heating of the sleeve heating device 5 when it rises. engage with it. When approaching the bottom, the inclination increases, and finally closes at the bottom, such as a circle of 3*3mm. The diameter of the bottom can be designed according to the size requirements of the hospital personnel for the embedding frame. The interface of the bottom circle is trapezoidal, so that the metal sheet 24 It is more convenient to embedding as an embedding frame. Preferably, a metal mesh is attached to the outside of the test tube 2, and the metal mesh can be formed by a metal sheet with good thermal conductivity or a small hole is opened on the metal sheet to facilitate the heating conduction of the sleeve heating device 5 and prevent the heating of the sleeve heating device 5. In the step, the paraffin in the test tube 2 is cooled and solidified; the inside of the test tube 2 wall of the test tube 2 has a symmetrically arranged first punching pipeline 21 and a second punching pipeline 22, and the symmetrical design can avoid the bias of the test tube 2 when it is centrifuged. The outlet end is set on the wall of the test tube 2 above the inner metal sheet 24 of the test tube 2, so that the liquid flow punched in during punching directly hits the area where the cell clumps are located, ensuring the breaking up of the cell clumps, and at the same time driving the inside of the test tube 2 from the bottom. The liquid flows and disperses evenly. The inlet end of the stamping pipeline is set on the outer side of the test tube 2 wall or the top of the test tube 2. It is preferably designed on the outer side of the test tube 2 wall to reduce the interference effect of the thickness of the test tube 2 wall. Design at the entrance end of the stamping pipeline The quick-plug interface 23, the quick-plug interface 23 can use the quick-plug method of the existing format structure, such as the self-locking type pressed by pressure, and the quick-plug interface 23 that is directly plugged and unplugged by pressing to unlock, etc., the structure is not limited ; There is a one-way valve in the stamping pipeline, the one-way valve can be a simple one-way cover plate, or a one-way valve structure of other structures, and the position is preferably at the inlet end and the outlet end to prevent the internal liquid from flowing out during centrifugation. Preferably, a liquid level detector is set to detect the liquid level in the test tube 2, and is controlled by the overall device. The type of the liquid level detector is not limited. For example, an infrared non-contact type is used, and it is arranged on the outside of the test tube 2, which is efficient and convenient.

The first stamping device 3 is used to connect the first stamping pipeline 21 and stamp reagents into the first stamping pipeline 21, including several reagent bottles 31, a first conduit, a first pressurizing pump, a solenoid valve, and a first mechanical arm, The first booster pump is installed on the first conduit pipeline, and the first conduit connects the reagents in several reagent bottles 31 respectively through the solenoid valve. A quick-plug interface 23 of a stamping pipeline 21, the first conduit can pressurize the reagent in the reagent bottle 31 into the first stamping pipeline 21 through the first pressurizing pump, the reagent bottle 31 includes a formulator for placing Reagent bottles 31 of liquid reagents such as forest, alcohol, eosin, and xylene, the corresponding conduits have multiple branches from the base to communicate with different reagent bottles 31, and each branch is provided with a solenoid valve to control the on-off of the connection. They are respectively used for stamping formalin, 95% alcohol, anhydrous alcohol, Yin Hong and xylene; the first mechanical arm and the second mechanical arm are similar in structure, arranged on both sides of the turntable 1, including the base and The retractable clamping part, the mechanical arm is a hollow structure, the pipelines of the first conduit and the second conduit pass through the cavity inside the mechanical arm, and are finally exposed to the clamping part, controlled by the internal controller. The layout of the test tube 2 is straight, the control design is simple, and the complicated control program design is avoided.

The second stamping device 4 is used to connect the second stamping pipeline 22 and stamp liquid paraffin in the second stamping pipeline 22, including a metal paraffin pool 41, a second conduit, a second pressurizing pump, a second mechanical arm, a second The booster pump is installed on the second conduit pipeline, one end of the second conduit is connected to the metal paraffin pool 41, the second mechanical arm controls the second conduit to connect with the quick-plug interface 23 of the second stamping pipeline 22, and the metal paraffin pool 41 is equipped with a heater , the heater is designed to heat the metal paraffin pool 41 in advance to obtain melted liquid paraffin before the liquid paraffin needs to be stamped in the preparation process. Preferably, an insulating layer is attached to the second conduit, and a relatively short distance between the metal paraffin pool 41 and the test tube 2 is designed to reduce the paraffin wax solidification by cooling.

The sleeve heating device 5 is arranged under the turntable 1 and can be controlled to move up and down. It includes a heating sleeve 51 that can be raised and lowered. The top of the heating sleeve 51 has a positioning protrusion 52 that is compatible with the positioning block 25 of the test tube 2, and can be selected according to needs. Appropriate sensors, such as contact type or infrared type, etc., when the heating sleeve 51 rises, the positioning protrusion 52 contacts the positioning block 25 of the test tube 2 to judge the rising stop, and the inner shape design of the heating sleeve 51 is consistent with the external shape of the test tube 2 Compatible with wrapping the bottom of the test tube 2, the heating sleeve 51 has a heating film 53, and meanwhile, the material of the heating sleeve 51 device is selected from a metal with good thermal conductivity, which is beneficial to heating and heat conduction. Preferably, the cooperation between the positioning block 25 and the positioning protrusion 52 can be omitted, and the fixed lifting distance can be used instead of the positioning block 25 or the protrusion of the quick-plug interface 23 can simplify the structure.

The liquid suction device 6 is used to absorb liquid from the test tube 2, including a collection bottle 61, a suction pipe, a working pump, a third mechanical arm, and the working pump is installed on the pipette pipeline, and the suction pipe communicates with the collection bottle 61, and the collection bottle 61 is used to collect the waste liquid drawn, including supernatant liquid or reagents, liquid paraffin, etc. that are not required according to the process requirements in the test tube 2. The third mechanical arm controls the suction pipe to extend into or out of the test tube 2. The third mechanical arm The arm is arranged on the top of the turntable 1, and has a base, a rotating part, and a retractable clamping part. The clamping part fixes the liquid suction pipe, and the rotating part is convenient to rotate to avoid the upper space during operation.

This embodiment also provides a stamping method for automatically preparing a cell wax block, comprising the steps of:

S1. Sampling, put the cell liquid of the cell wax block to be prepared into the test tube 2, and place the test tube 2 in the fixed hole 11 of the turntable 11 to fix; the cell liquid of the cell wax block to be prepared can be clinically extracted or collected by the operator Sampling, the sampling object can be human body fluid cells, such as pleural effusion, ascites, cervical exfoliated cells, serous cavity effusion, cerebrospinal fluid, sputum, etc.;

S2. form a cell mass, and gather dense substances at the bottom of the test tube 22 by centrifugation to form a cell mass;

Wherein, centrifugation means that the turntable 11 rotates at 1000-2000 rpm for 1-2 minutes, and then according to the process, the liquid suction device 6 sucks out the supernatant liquid or reagent and liquid paraffin in the test tube 2; The number of test tubes 2 is repeated multiple times in a row; the speed requirements and process duration during the process can be set by the operator according to the requirements, and the follow-up process is the same.

S3. Fixing, the solenoid valve is connected to the reagent for fixing, and the process of stamping reagent, natural precipitation, repeated stamping precipitation, and centrifugation is repeated. The process of this fixation lasts 1.5-2 hours. This process ensures that the reagent in the test tube 2 is higher than that at the bottom. Cell mass 5cm;

Among them, the reagent for fixation refers to neutral formalin with a concentration of 10%; the stamping reagent refers to the first stamping device 3 docking with the first stamping pipeline 21 and stamping the reagent into the first stamping pipeline 21 to drive the liquid in the test tube 2 flow, breaking up cell clumps, and breaking up can make all cells fully contact with the added reagents. If there is no above stamping process, the added reagents will only interact with the cells on the upper surface of the cell clumps deposited at the bottom of the test tube 2, while Unable to fully interact with the internal cells; natural precipitation refers to the natural precipitation of the cell liquid in test tube 2 for 5-15 minutes; repeated stamping precipitation refers to the process of repeating 2-4 times, stamping reagent/liquid paraffin, natural precipitation, repeated stamping can continue Adding new reagents can guarantee the fullness of the reaction, speed up the reaction time, and reduce the preparation time; natural precipitation refers to the natural precipitation of the cell liquid in the test tube 22 for 5-10 minutes; the purpose of adding and repeating the process by centrifugation is to control the concentration and liquid content of the internal reaction of old and new reagents. The height of the surface ensures sufficient reaction and improves the quality of preparation.

Preferably, when stamping reagents or liquid paraffin, the corresponding flow velocity requirement in the first conduit or the second conduit is 0.2-0.3m/s, so as to ensure the dispersion force and improve the preparation quality. For flow velocity detection, flow velocity detection sensors are installed on the pipelines of the first conduit and the second conduit, and further, when the pipelines are installed, they are installed near the ends of the conduits before the clamping part of the mechanical arm.

S4. Dehydration, including primary dehydration and secondary dehydration. For the primary dehydration, the solenoid valve communicates with the reagent for primary dehydration, and repeats the process of stamping reagents, natural precipitation, repeated stamping precipitation, and centrifugation. The primary dehydration process lasts 1- 1.5 hours; for the second dehydration, the solenoid valve is connected to the reagent for the second dehydration, and repeats the process of stamping reagents, natural precipitation, repeated stamping precipitation, and centrifugation. The secondary dehydration process lasts 1-1.5 hours;

Among them, the reagent for primary dehydration refers to alcohol with a concentration of 95%, and the reagent for secondary dehydration refers to absolute alcohol;

S5. Staining, the solenoid valve is connected to the reagent for dyeing, and the process of punching, natural precipitation, and repeated punching and precipitation is carried out. The dyeing process lasts 15-20 minutes. This process ensures that the reagent in the test tube 2 is 2 cm higher than the cell mass at the bottom;

Among them, the dyeing reagent refers to eosin, and the dyeing reagent can be selected according to actual needs, such as hematoxylin and eosin.

S6. Transparent, the electromagnetic valve is connected to the transparent reagent, and the process of stamping reagent, natural precipitation, repeated stamping precipitation, and centrifugation is repeated. The transparent process lasts 25-30 minutes. This process ensures that the reagent in the test tube 2 is higher than that at the bottom. Cell mass 5cm;

Among them, the reagent for transparency refers to xylene;

S7. Soak in paraffin, repeat the process of stamping liquid paraffin, heating test tube 2, natural precipitation, repeated stamping precipitation, test tube 2 detached from heating, and centrifugation. The toluene is replaced. During this process, the temperature of the test tube 2 is 60-70°C. This process ensures that the liquid paraffin in the test tube 2 is 5cm higher than the cell mass at the bottom;

Wherein, stamping liquid paraffin refers to that the second stamping device 4 is connected to the second stamping pipeline 22 and stamps liquid paraffin into the second stamping pipeline 22; while the heating of the test tube 2 refers to stamping the liquid paraffin, the sleeve heating device 5 controls the heating sleeve 51 rises until the heating sleeve 51 wraps the bottom of the test tube 2, and the heating film 53 in the heating sleeve 51 starts to heat; before the test tube 2 is separated from the heating finger and centrifuges, the sleeve heating device 5 controls the heating sleeve 51 to descend until it is separated from the test tube 2;

S8. After the above steps are completed, the cell mass has formed a wax block at the bottom of the test tube 2, and is located at the bottom. After the paraffin is cooled and solidified, the operator can take out the test tube 2, disassemble or cut and separate the bottom of the test tube 2 wall, and peel off the metal sheet 24 Obtain cell wax blocks for the next step of slicing.

Preferably, before punching the reagent/liquid paraffin, the liquid level height in the test tube 2 is detected by the liquid level detector, if the liquid level height is less than 2-3cm from the test tube 2 mouth distance, then the liquid suction device 6 will remove the reagent in the test tube 2 / Liquid paraffin is sucked out, and the height of the liquid in the test tube 2 is controlled to be 4-5cm lower than the mouth of the test tube 2, so as to avoid the overflow of the liquid surface when the test tube 2 is short.

The stamping-type automatic preparation of cell wax block and method thereof in this embodiment has a convenient, stable, smooth preparation process, strong applicability, is not affected by the size and quantity of cells to be prepared, and has low technical requirements for operators. And the heating energy consumption is low, and the new reagents added continuously by repeated punching can ensure the full reaction, and at the same time speed up the reaction time and reduce the preparation time.

The specific embodiments described herein are merely illustrative of the spirit of the invention. Those skilled in the art to which the present invention belongs can make various modifications or supplements to the described specific embodiments or adopt similar methods to replace them, but they will not deviate from the spirit of the present invention or go beyond the definition of the appended claims range.

Claims (8)

1. A stamping type device for preparing cell wax blocks is characterized by comprising a turntable, a test tube, a first stamping device, a second stamping device, a liquid suction device and a sleeve heating device;

the rotary disc is provided with a plurality of symmetrically arranged fixing holes for fixing the test tubes;

the test tube comprises a test tube body, a metal sheet is inlaid in the bottom of the test tube body, the surface of the metal sheet is smooth, the bottom of the test tube body where the metal sheet is located is of a detachable structure or can be cut and separated, a first stamping pipeline and a second stamping pipeline which are symmetrically arranged are arranged in the test tube wall of the test tube, the outlet end of the stamping pipeline is arranged on the test tube wall above the metal sheet in the test tube body, the inlet end of the stamping pipeline is arranged at the outer side part of the test tube wall or at the top of the test tube, the inlet end of the stamping pipeline is provided with a quick-plugging port, and a check valve is arranged in the stamping pipeline;

the first stamping device is used for butting a first stamping pipeline and stamping a reagent into the first stamping pipeline and comprises a plurality of reagent bottles, a first conduit, a first pressure pump, an electromagnetic valve and a first mechanical arm, wherein the first pressure pump is arranged on the first conduit, the first conduit is respectively communicated with the reagent in the plurality of reagent bottles through the electromagnetic valve, and the first mechanical arm controls the first conduit to be butted with a quick-plugging port of the first stamping pipeline;

the second stamping device is used for butting a second stamping pipeline and stamping liquid paraffin into the second stamping pipeline and comprises a metal paraffin pool, a second guide pipe, a second pressure pump and a second mechanical arm, wherein the second pressure pump is arranged on the second guide pipe, one end of the second guide pipe is communicated with the metal paraffin pool, and the second mechanical arm controls the second guide pipe to be butted with a quick-plugging port of the second stamping pipeline;

the liquid suction device is used for sucking liquid from a test tube and comprises a collecting bottle, a liquid suction pipe, a working pump and a third mechanical arm, wherein the working pump is arranged on a pipeline of the liquid suction pipe, the liquid suction pipe is communicated with the collecting bottle, and the third mechanical arm controls the liquid suction pipe to extend into or out of the test tube;

the sleeve heating device is arranged below the rotary table and used for heating the test tube and comprises a liftable heating sleeve, the inner shape of a heating sleeve container is matched with the outer shape of the test tube so as to wrap the bottom of the test tube, and a heating film is arranged in the heating sleeve;

there is also a liquid flow rate sensor for sensing the flow rate of liquid in the first and second conduits.

2. The apparatus for stamping cell wax pieces according to claim 1, wherein a metal mesh is attached to the outside of the test tube.

3. The device for stamping cell wax blocks according to claim 1, wherein the bottom of the test tube is in a circular truncated cone structure, and the test tube is reduced from the test tube mouth to the bottom circular truncated cone.

4. The device for stamping cell wax blocks according to claim 1, wherein the test tube has a positioning block at the middle lower part of the outer side, and the heating sleeve has a positioning protrusion at the top for fitting with the positioning block of the test tube.

5. The apparatus for stamping cell wax pieces according to claim 1, further comprising a liquid level detector for detecting the liquid level in the test tube.

6. A method for preparing cellular wax blocks based on a stamping device according to any one of claims 1 to 5, comprising the steps of:

s1, sampling: placing the cell sap of the cell wax block to be prepared into a test tube, and placing the test tube into a rotary table fixing hole for fixing;

s2, forming cell mass: gathering the dense substances at the bottom of the test tube by centrifugation to form cell masses;

wherein, the centrifugal rotating disc rotates for 1-2 minutes at 1000-2000 rpm, and the liquid absorbing device absorbs the supernatant liquid or reagent and liquid paraffin in the test tube;

s3, fixing: the electromagnetic valve is communicated with a fixing reagent to perform reagent stamping, natural precipitation, repeated stamping and precipitation and centrifugation, the fixing process lasts for 1.5-2 hours, and the process ensures that the reagent in the test tube is 5cm higher than the cell block mass at the bottom;

wherein the fixing reagent is neutral formalin with the concentration of 10%; the reagent stamping means that a first stamping device is in butt joint with a first stamping pipeline and stamps the reagent into the first stamping pipeline to drive liquid in the test tube to flow and break up cell blocks; natural precipitation means natural precipitation of cell liquid in a test tube for 5-15 minutes; the repeated stamping and precipitation refers to the process of stamping reagent or liquid paraffin and natural precipitation which are repeated for 2-4 times;

s4, dehydrating: the method comprises primary dehydration and secondary dehydration, wherein the primary dehydration is carried out by connecting an electromagnetic valve with a reagent for primary dehydration, stamping the reagent, naturally precipitating, repeatedly stamping and precipitating, and centrifuging, and the primary dehydration lasts for 1-1.5 hours;

the secondary dehydration process is that the electromagnetic valve is communicated with a reagent for secondary dehydration, the reagent is punched, naturally precipitated, repeatedly punched and precipitated and centrifuged, and the secondary dehydration process lasts for 1 to 1.5 hours;

wherein, the reagent for primary dehydration refers to alcohol with the concentration of 95 percent, and the reagent for secondary dehydration refers to absolute alcohol;

s5, dyeing: the electromagnetic valve is communicated with a reagent for dyeing, the stamping, the natural precipitation and the repeated stamping precipitation are carried out, the dyeing process lasts for 15-20 minutes, and the process ensures that the reagent in the test tube is 2cm higher than the cell block mass at the bottom;

wherein the staining reagent is eosin;

s6, transparency: the electromagnetic valve is communicated with a reagent for transparence, the reagent is punched, naturally precipitated, repeatedly punched and precipitated and centrifuged, the transparence process lasts for 25 to 30 minutes, and the process ensures that the reagent in the test tube is 5cm higher than the cell block mass at the bottom;

wherein, the reagent for transparence is dimethylbenzene;

s7, paraffin soaking: performing liquid paraffin stamping, test tube heating, natural precipitation, repeated stamping precipitation, test tube separation heating and centrifugation, wherein the paraffin soaking process lasts for 1-1.5 hours, the temperature of the test tube in the paraffin soaking process is 60-70 ℃, and the paraffin soaking process ensures that the liquid paraffin in the test tube is 5cm higher than the cell block mass at the bottom;

the liquid paraffin stamping device comprises a first stamping device, a second stamping device and a liquid paraffin stamping device, wherein the first stamping device is connected with a first stamping pipeline in a butt joint mode and stamps the liquid paraffin into the first stamping pipeline; when the test tube is heated, the sleeve heating device controls the heating sleeve to rise until the heating sleeve wraps the bottom of the test tube when the liquid paraffin is stamped, and a heating film in the heating sleeve starts to heat; before the test tube is separated from the heating, the sleeve heating device controls the heating sleeve to descend to be separated from the test tube;

s8, taking out the test tube, disassembling or cutting the bottom of the wall of the test tube, and stripping the metal sheet to obtain the cell wax block.

7. The method of claim 6, wherein repeating the stamping of the deposit further comprises the steps of: before stamping the reagent or the liquid paraffin, detecting the height of the liquid level in the test tube, if the distance between the liquid level and the opening of the test tube is less than 2-3cm, sucking the reagent or the liquid paraffin out of the test tube by the liquid sucking device, and controlling the height of the liquid in the test tube to be lower than the opening of the test tube by 4-5cm.

8. The method of claim 6, wherein the reagent or liquid paraffin is punched at a flow rate of 0.2 to 0.3m/s in the first conduit or the second conduit.

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