CN113390699B - Device and method for preparing cell wax block in stamping mode - Google Patents
Device and method for preparing cell wax block in stamping mode Download PDFInfo
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- CN113390699B CN113390699B CN202110562874.6A CN202110562874A CN113390699B CN 113390699 B CN113390699 B CN 113390699B CN 202110562874 A CN202110562874 A CN 202110562874A CN 113390699 B CN113390699 B CN 113390699B
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- 238000012360 testing method Methods 0.000 claims description 160
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- 239000012188 paraffin wax Substances 0.000 claims description 24
- 238000005119 centrifugation Methods 0.000 claims description 16
- 238000004043 dyeing Methods 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 12
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical group CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 11
- 239000001993 wax Substances 0.000 claims description 9
- 210000001503 joint Anatomy 0.000 claims description 7
- 230000001376 precipitating effect Effects 0.000 claims description 6
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- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical group [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 claims description 5
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- WDECIBYCCFPHNR-UHFFFAOYSA-N chrysene Chemical compound C1=CC=CC2=CC=C3C4=CC=CC=C4C=CC3=C21 WDECIBYCCFPHNR-UHFFFAOYSA-N 0.000 description 2
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/366—Moulds; Demoulding
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
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
Technical Field
The invention relates to the technical field of medical electronic instruments, in particular to a device and a method for preparing a cell wax block in a stamping mode.
Background
The cell wax block is prepared by extracting cells in liquid, fixing the cells in paraffin and preparing the fixed cells into a form similar to a tissue section, at present, a manual method is mostly adopted for preparing the cell wax block, the manual operation is time-consuming and labor-consuming, the prepared cell wax block is directly related to the experience level of operators, otherwise, the quality of the wax block is possibly uneven, and in addition, because reagents such as formalin, xylene and the like need to be added in the operation, the health of the operators can be harmed due to carelessness.
In the prior art, for example, a device and a method for automatically preparing a cell wax block are provided in a patent "CN 2014101669573" and a pressurized reaction device for preparing a cell wax block "in CN2014101643499, a series of steps such as fixing, dehydrating, transparentizing, dyeing, wax dipping, cooling and the like are automatically completed in an instrument, after the operation is finished, the cell block which is wax dipped can be formed in a test tube, the cell block is taken out and then placed in an embedding frame for embedding, and then the cell wax block can be sent to be sliced.
It has been found in practice that the apparatus and method also present certain problems and disadvantages. The problem is particularly pronounced when the body fluid contains a large number of cells. In operation, after the body fluid is poured into a test tube containing a semipermeable membrane, the reagent, moisture and small molecules in the body fluid penetrate through the semipermeable membrane, and the cells are retained on the semipermeable membrane. When a considerable amount of cells accumulate on the semi-permeable membrane, the filtration of liquids and small molecules is hindered, making the process slower and slower, and the process needs to be accelerated by applying negative pressure. The applied negative pressure is too small, and the effect is not obvious; oversize then can lead to the hole in the cell embedding semipermeable membrane slightly bigger, though through in vitro reverse application malleation can press the cell that partly blockked up the semipermeable membrane back again to the cell liquid in, still can have a small amount of cells that block up the semipermeable membrane to stay on the semipermeable membrane, through repeated operation like this many times, the filtration characteristic of last semipermeable membrane will disappear, leads to last effect not good. In addition, the paraffin wax is solidified when the temperature of the paraffin wax is lower than 40 ℃ in the wax dipping process, so that the test tube and the pressurizing and extracting device need to be kept at a certain temperature in the wax dipping process. The test tube is disposable and has few problems. However, the pressurizing and extracting device is operated circularly, and the paraffin adhered to the inner wall of the pressurizing and extracting device is accumulated more and more after the pressurizing and extracting device is operated for a long time. In order to ensure smooth work, the pressurizing and extracting devices need to be heated in the processes of other fixing, dehydrating, transparentizing, dyeing and the like, so that the operation is inconvenient and the energy consumption is overlarge.
In view of the above technical problems, it is desirable to improve.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a device and a method for preparing cell wax blocks in a stamping mode.
In order to achieve the purpose, the invention adopts the following technical scheme:
a punching device for preparing cell wax blocks comprises a rotary disc, a test tube, a first punching device, a second punching 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 the second stamping pipeline and stamping liquid paraffin into the second stamping pipeline and comprises a metal paraffin pool, a second conduit, a second pressure pump and a second mechanical arm, wherein the second pressure pump is arranged on the second conduit, one end of the second conduit is communicated with the metal paraffin pool, and the second mechanical arm controls the second conduit 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;
sleeve heating device sets up in the carousel below for heat the test tube, including the heating sleeve of liftable, the outside shape looks adaptation of shape and test tube in the heating sleeve container is in order to wrap up the test tube bottom, the heating film has in the heating sleeve.
Preferably, a metal net film is attached to the outer side of the test tube.
As the preferred scheme, the bottom of the test tube is of a round table structure, and the test tube is contracted from the test tube opening to the round table at the bottom.
As the preferred scheme, the lower part has the locating piece in the outside of test tube, heating sleeve top has the location arch with test tube locating piece looks adaptation.
Preferably, the apparatus for automatically preparing cell wax block further comprises a liquid level detector for detecting the liquid level in the test tube.
Preferably, the apparatus for automatically preparing a cellular wax block further comprises a liquid flow rate sensor for sensing a flow rate of the liquid in the first and second conduits.
A method for preparing a cell wax block by punching comprises the following steps:
s1, sampling, namely placing cell sap of a 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 a cell mass, and gathering a substance with high density at the bottom of the test tube through centrifugation to form the cell mass;
wherein, the centrifugal finger rotating disc rotates for 1-2 minutes at 1000-2000 rpm, and then the liquid absorbing device absorbs supernatant liquid or reagent and liquid paraffin in the test tube according to the process;
s3, fixing, wherein the electromagnetic valve is communicated with a fixing reagent, and repeated processes of reagent stamping, natural precipitation, repeated reagent stamping and precipitation and centrifugation are carried out, wherein 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 percent; 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/liquid paraffin and natural precipitation which is repeated for 2-4 times;
s4, dehydrating, including primary dehydrating and secondary dehydrating, wherein in the primary dehydrating, an electromagnetic valve is communicated with a reagent for primary dehydrating, and repeated processes of reagent stamping, natural precipitation, repeated stamping precipitation and centrifugation are carried out, and the primary dehydrating lasts for 1-1.5 hours; the secondary dehydration, wherein the electromagnetic valve is communicated with a reagent for secondary dehydration to perform repeated processes of reagent stamping, natural precipitation, repeated stamping precipitation and centrifugation, and the secondary dehydration process lasts for 1-1.5 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, wherein an electromagnetic valve is communicated with a reagent for dyeing, and the processes of stamping, natural precipitation and repeated stamping and precipitation are carried out, wherein 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, wherein the electromagnetic valve is communicated with a transparent reagent, repeated processes of reagent stamping, natural precipitation, repeated reagent stamping and precipitation and centrifugation are carried out, the transparency process lasts for 25-30 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, soaking paraffin, repeatedly punching liquid paraffin, heating a test tube, naturally precipitating, repeatedly punching and precipitating, separating the test tube from the heating and centrifuging, wherein the process lasts for 1-1.5 hours, the temperature of the test tube in the process is 60-70 ℃, and the process ensures that the liquid paraffin in the test tube is 5cm higher than a cell block mass at the bottom;
the liquid paraffin stamping device comprises a first stamping device, a second stamping device, a first stamping pipeline and a second stamping pipeline, wherein the first stamping device is connected with the first stamping pipeline in a butt joint mode; when the test tube is heated, the sleeve heating device controls the heating sleeve to ascend when the liquid paraffin is punched, until the heating sleeve wraps the bottom of the test tube, 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 until the heating sleeve is separated from the test tube;
s8, taking out the test tube, detaching or cutting the bottom of the test tube wall, and stripping the metal sheet to obtain the cell wax block.
Preferably, the process of repeatedly punching and depositing further comprises the following steps: before stamping the reagent/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/liquid paraffin out of the test tube by using a 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.
Preferably, the flow rate of the punching reagent or the liquid paraffin in the first conduit or the second conduit is 0.2 to 0.3m/s.
Compared with the prior art, the device and the method for preparing the cell wax block in the stamping mode have the advantages that the preparation process is convenient, stable and smooth, the applicability is strong, the device is not influenced by the size and the number of cells of cell sap to be prepared, the technical requirement on operators is low, the heating energy consumption is low, new reagents which are continuously added by repeatedly stamping can guarantee full reaction, the reaction time is shortened, and the preparation time is shortened.
Drawings
FIG. 1 is a schematic structural diagram of a stamping-type apparatus for automatically preparing cell wax blocks according to a first embodiment of the present invention;
FIG. 2 is a perspective view of the structure of a cuvette of the apparatus for stamping automatic preparation of cell wax blocks according to the first embodiment of the present invention;
FIG. 3 is a schematic front view showing the structure of a cuvette of the punch-type apparatus for automatically preparing cell wax pieces according to the first embodiment of the present invention;
FIG. 4 is a sectional view of the test tube of the punch-type apparatus for automatically preparing cell wax blocks according to the first embodiment of the present invention;
FIG. 5 is a schematic structural diagram of a heating sleeve of the punching-type apparatus for automatically preparing cell wax blocks according to the first embodiment of the present invention;
FIG. 6 is a schematic flow chart of a stamping-type apparatus for automatically preparing cell wax blocks according to a first embodiment of the present invention;
wherein: 1. a turntable; 11. a fixing hole; 2. a test tube; 21. a first stamping pipeline; 22. a second stamped pipe; 23. a quick-plugging port; 24. a metal sheet; 25. positioning a block; 3. a first punching device; 31. a reagent bottle; 4. a second stamping device; 41. a metal paraffin pool; 5. a sleeve heating device; 51. heating the sleeve; 52. positioning a projection; 53. heating the film; 6. a liquid suction device; 61. and (6) collecting the bottle.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It should be noted that the features in the following embodiments and examples may be combined with each other without conflict.
The first embodiment is as follows:
as shown in fig. 1 to 6, this embodiment proposes an apparatus for automatically preparing a cell wax block by stamping and a method thereof, in which the automatic operation of the steps of fixing a cell pellet, dehydrating, transparentizing, dyeing, waxing, cooling, etc. is performed in a designed test tube 2 without the need of a semi-permeable membrane in conjunction with positive and negative pressure.
Specifically, the device for automatically preparing the cell wax block comprises a rotary disc 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.
Wherein, carousel 1 has a plurality of symmetrical arrangements for fixed test tube 2's fixed orifices 11, carousel 1 can open 4 according to size and scale needs, or 8, 16 wait fixed orifices 11, once only can place corresponding quantity test tube 2 promptly, require the symmetry when placing to the biasing when preventing the centrifugation.
The bottom of the inner side of the test tube 2 is embedded with a metal sheet 24, the shape of the metal sheet 24 is attached to 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 the surface of the metal sheet, and after the metal sheet enters the stamping process, the cell mass can be scattered and cannot be attached to the metal sheet 24 all the time; 2 bottoms in test tube at 24 places of sheetmetal are detachable construction or can tailor the separation, detachable construction can be threaded connection, interference fit's grafting, the joint etc, can tailor the separation then the flexible material is chooseed for use to the 2 materials in test tube, for example plastics, it is unanimous with the test tube 2 of ordinary experiment, can tailor as disposable, can dismantle repeatedly usable as required, finish when the waxing step, treat test tube 2 internal paraffin cooling back, operating personnel can be convenient unpack test tube 2 apart, peel off sheetmetal 24, obtain the cell wax piece of preparation. The sheetmetal 24 can be designed to be directly embedded as an embedding frame, or transferred to the embedding frame as a transfer, and then slicing operation is carried out next step, meanwhile, when the sheetmetal 24 is heated in the preparation process, the local heat conduction at the bottom can be improved, the local preparation temperature requirement of the cell mass is stabilized, and the heat utilization efficiency is improved. The lower part has locating piece 25 in the outside of test tube 2 for with sleeve heating device 5 cooperation location, the adaptation design has the hole of dodging that supplies locating piece 25 to pass on carousel 1 fixed orifices 11. Further, 2 bottoms of test tube are designed into round platform structure, and test tube 2 is by 2 mouthfuls of necking down to the bottom round platform of test tube, and 2 oral area diameters of test tube are big, then constantly slope, the diameter reduces slightly downwards to can contact with it when convenient sleeve heating device 5 rises. The slope increases when arriving the bottom soon, receive for example 3 at the bottom in 3mm circular, its bottom diameter can be according to the size requirement design of hospital personnel to the embedding frame and decide, and bottom round platform interface is trapezoidal, makes 24 direct embedding as the embedding frame of sheetmetal more convenient. Preferably, a metal net film is attached to the outer side of the test tube 2, the metal net film can be formed by a metal sheet with good heat conductivity or a small hole formed in the metal sheet, so that the heating conduction of the sleeve heating device 5 is facilitated, and paraffin in the test tube 2 is prevented from being cooled and solidified in the wax dipping step; be provided with the check valve in the punching press pipeline, the check valve can be simple one-way cover plate, or the check valve structure of other structures, and the position is preferred in entry end, exit end, and inside liquid flows out when avoiding the centrifugation. Preferably, set up liquid level detector and be used for detecting liquid level in test tube 2, cooperation overall device control, liquid level detector type is not limited, for example chooses for use infrared contactless, sets up in the 2 outsides of test tube, and is high-efficient convenient.
The first punching device 3 is used for butting the first punching pipeline 21 and punching a reagent into the first punching pipeline 21, and comprises a plurality of reagent bottles 31, 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 pipeline, the first conduit is respectively communicated with the reagents in the plurality of reagent bottles 31 through the electromagnetic valve, the first mechanical arm comprises a base and a telescopic front clamping part, the first conduit is controlled to be butted with a quick-plugging port 23 of the first punching pipeline 21, the first conduit can press and punch the reagent in the reagent bottles 31 into the first punching pipeline 21 through the first pressure pump, the reagent bottles 31 comprise reagent bottles 31 for placing liquid reagents such as formalin, alcohol, eosin branches and xylene, the corresponding conduits are provided with a plurality of reagent bottles 31 from the base and are respectively communicated with different reagent bottles 31, each branch is provided with an electromagnetic valve for controlling the connection and disconnection of the connection and disconnection, so as to be respectively used for punching formalin, alcohol with the concentration of 95%, absolute alcohol, chrysene, xylene and the like; the first mechanical arm and the second mechanical arm are similar in structure and are arranged on two sides of the turntable 1, the mechanical arm is of a hollow structure and is provided with a cavity passing through the inner part of the mechanical arm, and the first conduit and the second conduit are finally exposed out of the clamping part and are controlled by an internal controller to stretch and retract, the two sides of the arrangement straight test tube 2 are simple in control design, and complicated control program design is avoided.
Second stamping device 4 is used for butt joint second punching press pipeline 22 and to punching press liquid paraffin in the second punching press pipeline 22, including metal paraffin pond 41, the second pipe, the second force (forcing) pump, the second arm, the second force (forcing) pump is installed on the second pipe pipeline, second pipe one end intercommunication metal paraffin pond 41, second arm control second pipe butt joint second punching press pipeline 22's fast interface 23, metal paraffin pond 41 is with the heater, the heater design is before preparation process needs punching press liquid paraffin, just heat in advance in order to obtain the liquid paraffin that melts to metal paraffin pond 41. Preferably, an insulating layer is attached to the second guide pipe, and a closer distance between the metal paraffin pool 41 and the test tube 2 is designed to reduce the paraffin freezing.
Sleeve heating device 5 sets up in carousel 1 below, can be controlled and reciprocate, heating sleeve 51 including the liftable, heating sleeve 51 top has the protruding 52 in location with 2 locating piece 25 looks adaptations of test tube, select suitable sensor as required, for example contact or infrared formula etc., protruding 52 in location and the contact of 2 locating pieces of test tube 25 can judge to rise and stop when heating sleeve 51 rises, shape design and 2 outside shape looks adaptations of test tube are in order to wrap up 2 bottoms of test tube in the heating sleeve 51 container, heating film 53 has in the heating sleeve 51, and simultaneously, the good metal of thermal conductivity is selected for use to the material of heating sleeve 51 device, do benefit to the heating, heat conduction. Preferably, the cooperation of the positioning block 25 and the positioning protrusion 52 may be omitted, and the lifting fixing path may replace the positioning block 25 or the protrusion of the quick-insertion port 23 may replace the positioning block 25, thereby simplifying the structure.
The embodiment also provides a method for stamping the device for automatically preparing the cell wax block, which comprises the following steps:
s1, sampling, namely putting cell sap of a cell wax block to be prepared into a test tube 2, and putting the test tube 2 into a fixing hole 11 of a rotary disc 11 for fixing; the cell fluid of the cell wax block to be prepared can be clinically extracted or collected and sampled in a centralized way by an operator, and the sampled object can be human body fluid cells, such as pleural effusion, ascites, cervical exfoliated cells, serosal cavity effusion, cerebrospinal fluid, sputum and the like;
s2, forming a cell mass, and gathering substances with high density at the bottom of the test tube 22 through centrifugation to form the cell mass;
wherein, the centrifugal finger rotating disc 11 rotates for 1-2 minutes at 1000-2000 rpm, and then the liquid absorbing device 6 absorbs the supernatant liquid or reagent and liquid paraffin in the test tube 2 according to the process; in the process, repeated operation is continuously carried out for a plurality of times according to the number of the test tubes 2 on the turntable 1; the rotating speed requirement and the process duration in the process can be set by an operator according to the requirement, and the subsequent processes are the same.
S3, fixing, wherein the electromagnetic valve is communicated with a fixing reagent, and repeated processes of reagent stamping, natural precipitation, repeated reagent stamping and precipitation and centrifugation are carried out, wherein the fixing process lasts for 1.5-2 hours, and the process ensures that the reagent in the test tube 2 is 5cm higher than the cell block mass at the bottom;
wherein the fixing reagent is neutral formalin with the concentration of 10 percent; the stamping reagent means that the first stamping device 3 is butted with the first stamping pipeline 21 and stamps the reagent into the first stamping pipeline 21 to drive the liquid in the test tube 2 to flow, so as to break up cell agglomerates, all cells can be fully contacted with the added reagent, and if the stamping process is not available, the added reagent only can act on the cells deposited on the upper surface of the cell agglomerates at the bottom of the test tube 2 and cannot fully act on the cells inside; natural precipitation means natural precipitation of cell liquid in the test tube 2 for 5-15 minutes; the repeated stamping and precipitation refers to the process of stamping the reagent/liquid paraffin and natural precipitation which is repeated for 2-4 times, and new reagents can be continuously added by repeated stamping, so that the full reaction can be ensured, the reaction time is shortened, and the preparation time is shortened; natural precipitation refers to natural precipitation of cell liquid in the test tube 22 for 5-10 minutes; the aim of the repeated process of centrifugal addition is to control the reaction concentration and the liquid level height of new and old reagents in the reaction system, ensure the full reaction and improve the preparation quality.
Preferably, when the reagent or the liquid paraffin is punched, the flow speed requirement corresponding to the inside of the first conduit or the second conduit is 0.2-0.3m/s, the breaking force is guaranteed, the preparation quality is improved, the flow speed can be controlled through the flow speed detection in the pressure pump, or flow speed detection sensors are arranged on the pipelines of the first conduit and the second conduit, and further, when the pipelines are arranged, the flow speed detection sensors are arranged near the end part of the conduit in front of the mechanical arm clamping part.
S4, dehydrating, including primary dehydration and secondary dehydration, wherein the primary dehydration is performed by connecting the magnetic valve with a reagent for primary dehydration and performing repeated processes of reagent stamping, natural precipitation, repeated stamping precipitation and centrifugation, and the primary dehydration lasts for 1-1.5 hours; the secondary dehydration, wherein the electromagnetic valve is communicated with a reagent for secondary dehydration to perform repeated processes of reagent stamping, natural precipitation, repeated stamping precipitation and centrifugation, and the secondary dehydration process lasts for 1-1.5 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, wherein an electromagnetic valve is communicated with a reagent for dyeing, and the processes of stamping, natural precipitation and repeated stamping and precipitation are carried out, wherein the dyeing process lasts for 15-20 minutes, and the process ensures that the reagent in the test tube 2 is 2cm higher than the cell block mass at the bottom;
the reagent for dyeing is eosin, the reagent for dyeing can be selected according to actual needs, such as hematoxylin, the requirement of an eosin dyeing process is simple and convenient, centrifugation is not needed in the scheme, and the process time is saved;
s6, transparency, wherein an electromagnetic valve is communicated with a transparent reagent, repeated processes of reagent stamping, natural sedimentation, repeated reagent stamping and centrifugation are carried out, the transparency process lasts for 25-30 minutes, and the process ensures that the reagent in the test tube 2 is 5cm higher than the cell block mass at the bottom;
wherein, the reagent for transparence is dimethylbenzene;
s7, soaking paraffin, repeatedly punching liquid paraffin, heating the test tube 2, naturally precipitating, repeatedly punching and precipitating, removing the test tube 2 from heating and centrifuging, wherein the process lasts for 1-1.5 hours, residual dimethylbenzene on the cells after the cells are subjected to the transparentization treatment is replaced, the temperature of the test tube 2 is 60-70 ℃ in the process, and the process ensures that the liquid paraffin in the test tube 2 is 5cm higher than a cell block mass at the bottom;
the liquid paraffin stamping means that the second stamping device 4 is in butt joint with the second stamping pipeline 22 and stamps the liquid paraffin into the second stamping pipeline 22; when the test tube 2 is heated, namely liquid paraffin is punched, the sleeve heating device 5 controls the heating sleeve 51 to ascend 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, the sleeve heating device 5 controls the heating sleeve 51 to descend until the test tube 2 is separated;
s8, after the steps are finished, the cell block masses form wax blocks at the bottoms of the test tubes 2 and are positioned at the bottommost parts, after paraffin is cooled and solidified, an operator can take out the test tubes 2, the bottoms of the walls of the test tubes 2 are detached or cut, the metal sheets 24 are peeled off to obtain the cell wax blocks, and the next slicing operation is carried out.
Preferentially, before punching press reagent/liquid paraffin, detect the liquid level height in the test tube 2 through liquid level height detector, if the liquid level height apart from 2 mouthfuls of test tube apart from being less than 2-3cm, then imbibition device 6 is less than 2 mouthfuls of test tube 2 reagent/liquid paraffin suction, and the liquid height in the control test tube 2 is less than 2 mouthfuls of test tube 4-5cm, avoids overflowing because of the liquid level of chooseing for use test tube 2 when short, and the concentration of new reagent when the imbibition can improve the reaction simultaneously for reaction time.
The device and the method for automatically preparing the cell wax block by stamping have the advantages of convenient, stable and smooth preparation process, strong applicability, no influence of the size and the number of the cells of the cell sap to be prepared, low technical requirement on operators, low heating energy consumption, capability of ensuring full reaction by repeatedly stamping new reagents which are continuously added, accelerated reaction time and shortened preparation time.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
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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| CN212180405U (en) * | 2020-06-16 | 2020-12-18 | 湖北医药学院 | Device for automatically preparing cell wax block |
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| US4656047A (en) * | 1986-03-06 | 1987-04-07 | Kok Lanbrecht P | Rapid method for cell block preparation |
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| RU2740431C1 (en) * | 2020-03-12 | 2021-01-14 | Федеральное государственное автономное образовательное учреждение высшего образования "Балтийский федеральный университет имени Иммануила Канта" (БФУ им. И. Канта) | Method of preparing cell blocks based on cervical exfoliative material and cervical canal |
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