CN113390700B - Automatic cell wax block preparation system and method based on centrifugal method - Google Patents
Automatic cell wax block preparation system and method based on centrifugal method Download PDFInfo
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- G—PHYSICS
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- 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
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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
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- 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
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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
- G01N1/44—Sample treatment involving radiation, e.g. heat
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Abstract
The invention relates to the technical field of medical electronic instruments, in particular to an automatic cell wax block preparation system based on a centrifugal method and a method thereof. The automatic preparation system and the method for the cell wax block based on the centrifugal method have the advantages of convenient, stable and smooth preparation process, strong applicability, no influence by the size and the number of the cells of the cell sap to be prepared, low heating energy consumption, low requirement on the installation precision of the device and low technical requirement on operators.
Description
Technical Field
The invention relates to the technical field of medical electronic instruments, in particular to a system and a method for automatically preparing a cell wax block based on a centrifugal method.
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 that the apparatus and method also present problems and disadvantages in practice. 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 have accumulated 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 a 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 provide a system and a method for automatically preparing a cell wax block based on a centrifugal method, aiming at the defects of the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
an automatic cell wax block preparation system based on a centrifugal method comprises a rotary table, a test tube, a paraffin extraction device, a liquid taking device, a sleeve heating device and a stirring 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 wall, a positioning block is arranged at the middle lower part of the outer side of the test tube wall, a metal sheet is embedded at the bottom of the inner side of the test tube wall, the surface of the metal sheet is smooth, and the bottom of the test tube wall where the metal sheet is located is of a detachable structure or can be cut and separated;
the paraffin extraction device is used for adding or absorbing liquid paraffin into the test tube;
the liquid taking device is used for adding a reagent into the test tube and taking liquid from the test tube;
the sleeve heating device is arranged below the turntable and comprises a liftable heating sleeve, a positioning protrusion matched with a positioning block of the test tube wall is arranged at the top of the heating sleeve, the inner shape of the 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;
the stirring device comprises a stirring arm and a stirring rod, wherein the stirring arm controls the stirring rod to extend into or out of the test tube and controls the stirring action.
Preferably, a metal net film is attached to the outer side of the tube wall.
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 preferred scheme, the paraffin extraction device includes that first arm, paraffin absorb pipeline, first working pump, first solenoid valve, metal paraffin pond is with the heater, first arm control paraffin absorbs the pipeline and stretches into or stretch out the test tube, paraffin absorbs pipeline intercommunication metal paraffin pond, first working pump, first solenoid valve are installed on paraffin absorption pipeline.
As a preferred scheme, get liquid device and include second arm, pipe, second working pump, second solenoid valve, a plurality of reagent bottle, second arm control pipe stretches into or stretches out the test tube, the pipe communicates a plurality of reagent bottles, second working pump, second solenoid valve are installed on the pipe.
A method of a cell wax block automatic preparation system based on a centrifugal method 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 revolutions per second, and then the liquid taking device sucks out supernatant liquid or reagent in the test tube or the paraffin extracting device sucks out liquid paraffin on the upper layer according to the process;
s3, fixing, namely repeatedly adding a fixing reagent, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the fixing process lasts for 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, adding fixing reagent refers to adding 5-10ml neutral formalin with concentration of 10% into the test tube; the stirring device controls the stirring rod to extend into the test tube for stirring, and the cell blocks extend out of the stirring rod after being stirred and scattered; natural precipitation refers to natural precipitation of cell liquid in a test tube for 5-10 minutes; the repeated stirring and precipitation refers to the process of stirring and natural precipitation repeated for 3-4 times;
s4, dehydrating, namely primary dehydrating and secondary dehydrating, wherein the primary dehydrating is repeated, a reagent for primary dehydrating is added, stirring, natural precipitation, repeated stirring precipitation and centrifugation are carried out, and the primary dehydrating lasts for 1.5 hours; the secondary dehydration is carried out repeatedly, and the processes of adding a reagent for secondary dehydration, stirring, natural precipitation, repeated stirring, precipitation and centrifugation are carried out, wherein the secondary dehydration lasts for 1.5 hours;
wherein, adding reagent for primary dehydration to add 95% alcohol into the test tube; adding a reagent liquid-taking device for secondary dehydration to add absolute alcohol into the test tube;
s5, dyeing, namely adding a reagent for dyeing, stirring, naturally precipitating, and repeatedly stirring and precipitating, wherein the dyeing process lasts for 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, adding a dyeing reagent refers to adding 1 ml-2 ml eosin into a test tube by a liquid extraction device;
s6, performing repeated processes of adding a reagent for transparency, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the transparent process lasts for 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, adding a transparent reagent-indicating liquid device to add dimethylbenzene into the test tube;
s7, soaking paraffin, repeating the processes of adding liquid paraffin, heating a test tube, stirring, naturally precipitating, repeatedly stirring and precipitating, separating the test tube from heating and centrifuging, wherein the process lasts for 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;
wherein, adding the liquid paraffin means adding the liquid paraffin into the test tube by a paraffin extracting device; when the liquid paraffin is added in the test tube heating, the sleeve heating device controls the heating sleeve to ascend until the positioning protrusion of the heating sleeve abuts against the positioning block of the test tube wall, the heating sleeve wraps the bottom of the test tube at the moment, 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 test tube is separated from the heating sleeve;
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.
Preferably, the pipeline of the liquid taking device in the step is communicated with clean water, and the pipeline of the liquid taking device is flushed by the clean water before the reagent is added into the test tube and before the liquid is taken from the test tube.
Compared with the prior art, the automatic cell wax block preparation system and method based on the centrifugal method 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 heating energy consumption, low requirement on installation precision of the device and low technical requirement on operators.
Drawings
FIG. 1 is a schematic structural diagram of a system for automatically preparing cell wax blocks by centrifugation according to a first embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a test tube of a system for automatically preparing cell wax blocks based on centrifugation according to a first embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a heating sleeve of a system for automatically preparing cell wax blocks based on a centrifugation method according to a first embodiment of the present invention;
FIG. 4 is a schematic flow chart of a method for automatically preparing a cellular wax block system based on centrifugation according to a first embodiment of the present invention;
wherein: 1. a turntable; 11. a fixing hole; 2. a test tube; 21. the wall of the test tube; 22. positioning blocks; 23. a metal mesh film; 24. a metal sheet; 3. a paraffin extraction device; 31. a metal paraffin pool; 4. a liquid taking device; 41. a reagent bottle; 5, sleeve heating device; 51. heating the sleeve; 52. positioning the projection; 53. heating the film; 6. a stirring device; 61. a stirring arm; 62. a stirring rod.
Detailed Description
The following embodiments of the present invention are provided by way of specific examples, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. 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 is to 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 FIGS. 1 to 4, this embodiment is based on the centrifugation method, and proposes an apparatus and a method for automatically preparing a cell wax pellet by centrifugation, in which the automatic operations of fixing, dehydrating, clearing, staining, waxing, cooling, etc. of the cell pellet are performed in a designed test tube 2 without using a pressurizing and extracting apparatus.
Specifically, the device for automatically preparing the cell wax block comprises a rotary disc 1, a test tube 2, a paraffin extracting device 3, a liquid taking device 4, a sleeve heating device 5 and a stirring device 6.
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 test tube 2 comprises a test tube wall 21, a metal sheet 24 is embedded at the bottom of the inner side of the test tube wall 21, 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 and the cell mass enter the stirring process at the same time, the cell mass can be scattered and cannot be attached to the metal sheet 24 all the time; the test tube wall 21 bottom at 24 places of sheetmetal is 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 test tube wall 21 material chooses for use flexible material, for example plastics, it is unanimous with 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 interior 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 design directly as the embedding frame embedding, perhaps transfer to the embedding frame as the transfer in, carry out slicing operation next step, and sheetmetal 24 can promote the local heat-conduction in bottom when the preparation process heating simultaneously, stabilizes the local preparation temperature requirement of cell piece mass, promotes hot availability factor. The middle lower part of the outer side of the test tube wall 21 is provided with a positioning block 22 for matching and positioning with the sleeve heating device 5, and an avoidance hole for the positioning block 22 to pass through is designed on the fixing hole 11 of the rotary table 1 in a matching way. Further, the bottom of the test tube 2 is designed into a circular truncated cone structure, the test tube 2 is contracted from the opening of the test tube 2 to the circular truncated cone at the bottom, the diameter of the opening of the test tube wall 21 is large, and then the test tube wall is inclined downwards continuously and the diameter is slightly reduced, so that the sleeve heating device 5 can be contacted with the sleeve heating device when ascending. 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 23 is attached to the outer side of the test tube wall 21, and the metal net film 23 may be formed by a metal sheet with good thermal conductivity or small holes formed on the metal sheet, so as to facilitate the heating conduction of the sleeve heating device 5 and prevent the paraffin in the test tube 2 from cooling and solidifying in the paraffin dipping step.
Extracting paraffin device 3 is used for adding or absorbing liquid paraffin to test tube 2, including first arm, the paraffin absorbs the pipeline, a work pump, a solenoid valve, metal paraffin pond 31 is with the heater, the heater design is before liquid paraffin need be added in the preparation process, just heat in advance in order to acquire the liquid paraffin that melts metal paraffin pond 31, first arm control paraffin absorbs the pipeline and stretches into or stretch out test tube 2, paraffin absorbs pipeline intercommunication metal paraffin pond 31, a work pump, a solenoid valve is installed on paraffin absorbs the pipeline, a motion for control switch and absorption or addition. Preferably, absorb the attached heat preservation of pipeline at paraffin, design the nearer distance between metal paraffin pond 31 and test tube 2 simultaneously, reduce paraffin and receive the cold solidification. Further, metal paraffin pond 31 design has two branch ponds, and one of them is as the collecting pit of the old liquid paraffin that uses, and simultaneously in order to reduce cross contamination, the design is absorb and is drawn the pipeline with the paraffin that adds and be the double-circuit, and this double-circuit laminates each other, can absorb the heat each other, and the temperature that paraffin kept stable liquid state needs in the stable preparation process.
The liquid taking device 4 is used for adding a reagent into the test tube 2 and taking a liquid from the test tube 2, and comprises a second mechanical arm, a conduit, a second working pump, a second electromagnetic valve and a plurality of reagent bottles 41, wherein the second mechanical arm controls the conduit to extend into or out of the test tube 2, the conduit is communicated with the plurality of reagent bottles 41, the second working pump and the second electromagnetic valve are installed on the conduit, the conduit can suck the reagent in the reagent bottles 41 and add the reagent into the test tube 2 or take the liquid from the test tube 2 into the reagent bottles 41, each reagent bottle 41 comprises the reagent bottles 41 for containing liquid reagents such as formalin, alcohol, eosin and xylene, and the corresponding conduit is provided with a plurality of branches from a base and is respectively communicated with different reagent bottles 41 so as to respectively suck the formalin, the alcohol with the concentration of 95%, the absolute alcohol, the eosin and the xylene; preferably, the reagent bottles 41 further include a clear water reagent bottle 41 for placing cleaning, a waste liquid bottle for collecting waste liquid, and a reagent bottle 41 for pumping cell liquid to be prepared, the former can reduce cross contamination, the latter can replace pouring operation and further batch automation as required; the first mechanical arm and the second mechanical arm are similar in structure and comprise a base, a rotating part and a clamping part, the base is provided with a plurality of through holes, the mechanical arm is of a hollow structure, a controller is arranged in the rotating part, and a pipeline of a catheter or paraffin suction pipeline is arranged in a cavity in the mechanical arm and finally exposed out of the clamping part.
Agitating unit 6 includes stirring arm 61, stirring rod 62, and stirring arm 61 control stirring rod 62 stretches into or stretches out test tube 2, and control stirring action, and stirring arm 61 is including rotation portion and clamping part, and stirring rod 62 is located clamping part department, and stirring rod 62's tip design has detachable stirring head, can select different stirring heads according to the container of stirring and stirring dynamics to stir bottom sedimentary cell lump, break up when being convenient for stir.
The embodiment also provides a method for automatically preparing a cell wax block by a centrifugal method, 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 turntable 1 for fixing; the cell sap of the cell wax block to be prepared can be clinically extracted or collected and sampled in a centralized way by an operator, the sampled object can be the cells of body fluid of a human body, such as pleural effusion, ascites, cervical exfoliated cells, serosal cavity effusion, cerebrospinal fluid, sputum and the like, and the pouring operation can also be designed and finished by the liquid taking device 4;
s2, forming a cell mass, and gathering substances with high density at the bottom of the test tube 2 through centrifugation to form the cell mass;
wherein, the centrifugal finger rotary disc 1 rotates for 1-2 minutes at 1000-2000 revolutions per second, and then the liquid taking device 4 sucks out supernatant liquid or reagent in the test tube 2 or the paraffin extracting device 3 sucks out liquid paraffin on the upper layer according to the process; the liquid taking process or the process of extracting the upper-layer liquid paraffin is finished by controlling the pipeline of the clamping part to extend into or extend out of the test tube 2 by the corresponding mechanical arm; in the process, the test tubes 2 on the turntable 1 are continuously carried out for a plurality of times, preferably, the test tubes are washed by clean water before the test tubes 2 are sucked each time and then added with reagents, so that the cross contamination of samples is prevented; if the color of the upper liquid of the test tube 2 is turbid, clear water can be sucked into the test tube 2, the supernatant is sucked after centrifugation, the operator can set the number of times of repetition according to the situation, or photoelectric detection and turbidity detection are designed to automatically judge and execute repetition.
S3, fixing, namely repeatedly adding a fixing reagent, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the fixing process lasts for 2 hours, the time can be set by an operator as required, and the process ensures that the reagent in the test tube 2 is 5cm higher than the cell block mass at the bottom;
wherein, a reagent for fixing is added, a liquid-taking device 4 is used for adding 5-10ml of 10 percent neutral formalin into the test tube 2; the stirring means that the stirring device 6 controls the stirring rod 62 to extend into the test tube 2 for stirring action, the cell mass block is stirred and scattered and then extends out of the stirring rod 62, all cells can be fully contacted with the added reagent, and if the stirring process is not available, the added reagent only can act on the cells deposited on the upper surface of the cell mass block at the bottom of the test tube 2 but cannot fully act on the cells inside; natural precipitation means natural precipitation of cell liquid in the test tube 2 for 5-10 minutes; the repeated stirring and precipitation refers to the process of stirring and natural precipitation repeated for 3-4 times; the repeated centrifugal adding process aims to prevent the performance of the acted reagent from changing, the repeated centrifugal process and the new reagent adding are designed in the extraction process, the full reaction is ensured, and the preparation quality is improved.
S4, dehydrating, including primary dehydration and secondary dehydration, wherein the primary dehydration is repeated, a reagent for primary dehydration is added, and the processes of stirring, natural precipitation, repeated stirring precipitation and centrifugation are carried out, so that the cells are fully dehydrated, the primary dehydration lasts for 1.5 hours, and the time can be set by an operator as required; the secondary dehydration is carried out repeatedly, the processes of adding a reagent for secondary dehydration, stirring, natural precipitation, repeated stirring precipitation and centrifugation are carried out, the primary dehydration process is repeated, the secondary dehydration process lasts for 1.5 hours, and the time can be set by an operator according to the requirement;
wherein, a reagent extracting device 4 for primary dehydration is added to add 95 percent alcohol into the test tube 2; adding a reagent extracting device 4 for secondary dehydration to add absolute alcohol into the test tube 2;
s5, dyeing, namely adding a reagent for dyeing, stirring, naturally precipitating, and repeatedly stirring and precipitating, wherein the dyeing process lasts for 20 minutes, the time can be set by an operator according to needs, and the process ensures that the reagent in the test tube 2 is 2cm higher than the cell mass block at the bottom;
wherein, adding 1 ml-2 ml eosin to the test tube 2 with the reagent for dyeing instruction liquid device 4, the dyeing reagent can be selected according to the actual need, such as hematoxylin, the eosin dyeing process is simple and convenient, the scheme can not need centrifugation, the process time is saved;
s6, performing repeated processes of adding a reagent for transparency, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the transparent process lasts for 30 minutes, the time can be set by an operator according to needs, and the process ensures that the reagent in the test tube 2 is 5cm higher than the cell block mass at the bottom;
wherein, a reagent extracting liquid device 4 for transparency is added to add dimethylbenzene into the test tube 2;
s7, soaking paraffin, repeatedly adding liquid paraffin, heating the test tube 2, stirring, naturally precipitating, repeatedly stirring and precipitating, removing the test tube 2 from heating and centrifuging, wherein the process lasts for 1.5 hours, residual dimethylbenzene on the cells after the cell is subjected to the transparentization treatment is replaced, the time can be set by an operator according to needs, 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;
wherein, adding the liquid paraffin means that the paraffin extracting device 3 melts the paraffin in the paraffin pool, and the melted paraffin is added into the test tube 2 and then the liquid paraffin is added into the test tube 2; when test tube 2 heating indicates adding liquid paraffin, sleeve heating device 5 control heating sleeve 51 rose, stopped when heating sleeve 51 location arch 52 leaned on with test tube wall 21 locating piece 22, heating sleeve 51 parcel test tube 2 bottom this moment, heating film 53 begins the heating in the heating sleeve 51, has heating film 53 in the sleeve to guarantee that the paraffin in the test tube 2 does not solidify because of receiving the cold; before the test tube 2 is heated in a breaking-away mode, the sleeve heating device 5 controls the heating sleeve 51 to descend until the heating sleeve is separated from the test tube 2, the centrifugal turntable 1 stops rotating and then ascends again according to process requirements, after paraffin in the test tube 2 is melted, old liquid paraffin is sucked out, and new liquid paraffin is added;
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, disassemble or cut the bottoms of the separated test tube walls 21, peel off the metal sheets 24 to obtain the cell wax blocks, and perform the next slicing operation.
The system and the method for automatically preparing the cell wax block based on the centrifugal method 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 heating energy consumption, low requirement on the installation precision of the device and low technical requirement on operators.
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 (4)
1. An automatic cell wax block preparation system based on a centrifugal method is characterized by comprising a turntable, a test tube, a paraffin extraction device, a liquid taking device, a sleeve heating device and a stirring 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 wall, wherein a positioning block is arranged at the middle lower part of the outer side of the test tube wall, a metal sheet is embedded at the bottom of the inner side of the test tube wall, the surface of the metal sheet is smooth, and the bottom of the test tube wall where the metal sheet is positioned is of a detachable structure or can be cut and separated;
the paraffin extracting device is used for adding or sucking liquid paraffin into the test tube;
the liquid taking device is used for adding a reagent into the test tube and taking liquid from the test tube;
the sleeve heating device is arranged below the turntable and comprises a liftable heating sleeve, a positioning protrusion matched with a positioning block of the test tube wall is arranged at the top of the heating sleeve, the inner shape of the 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;
the stirring device comprises a stirring arm and a stirring rod, wherein the stirring arm controls the stirring rod to extend into or out of the test tube and controls the stirring action;
the bottom of the test tube is of a round table structure, and the test tube is contracted from a test tube opening to a round table at the bottom;
the paraffin extraction device comprises a first mechanical arm, a paraffin suction pipeline, a first working pump, a first electromagnetic valve and a metal paraffin pool, wherein a heater is attached to the metal paraffin pool, the first mechanical arm controls the paraffin suction pipeline to extend into or out of the test tube, the paraffin suction pipeline is communicated with the metal paraffin pool, and the first working pump and the first electromagnetic valve are installed on the paraffin suction pipeline;
get liquid device and include second arm, pipe, second work pump, second solenoid valve, a plurality of reagent bottle, second arm control pipe stretches into or stretches out the test tube, a plurality of reagent bottles of pipe intercommunication, second work pump, second solenoid valve are installed on the pipe.
2. The system for automatically preparing cell wax by centrifugation according to claim 1, wherein a metal net film is attached to the outer side of the test tube wall.
3. A method based on a centrifugation-based automatic cellular wax block preparation system according to any one of claims 1-2, 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 a 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, the liquid taking device sucks out supernatant liquid or reagent in the test tube, or the paraffin extracting device sucks out liquid paraffin on the upper layer;
s3, fixing: adding a fixing reagent, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the fixing process lasts for 2 hours, and the fixing process ensures that the reagent in the test tube is 5cm higher than the cell block mass at the bottom;
wherein, adding fixing reagent refers to adding 5-10ml of 10% neutral formalin into the test tube;
the stirring device controls the stirring rod to extend into the test tube for stirring, and the cell block masses extend out of the stirring rod after being stirred and scattered; natural precipitation means natural precipitation of cell liquid in a test tube for 5-10 minutes; the repeated stirring and precipitation refers to repeated stirring and natural precipitation for 3-4 times;
s4, dehydration: the method comprises primary dehydration and secondary dehydration, wherein the primary dehydration comprises the steps of adding a reagent for primary dehydration, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, and the primary dehydration lasts for 1.5 hours; the secondary dehydration process comprises the steps of adding a reagent for secondary dehydration, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the secondary dehydration process lasts for 1.5 hours;
wherein, adding a reagent for primary dehydration, namely adding alcohol with the concentration of 95 percent into a test tube by a liquid extraction device; adding a reagent liquid-taking device for secondary dehydration to add absolute alcohol into the test tube;
s5, dyeing: adding a reagent for dyeing, stirring, naturally precipitating, and repeatedly stirring and precipitating, wherein the dyeing process lasts for 20 minutes, and the dyeing process ensures that the reagent in the test tube is 2cm higher than the cell mass at the bottom;
wherein, adding a reagent for dyeing, namely adding 1-2 ml of eosin into a test tube by using a liquid extraction device;
s6, transparency: adding a reagent for transparence, stirring, naturally precipitating, repeatedly stirring, precipitating and centrifuging, wherein the transparence process lasts for 30 minutes, and the reagent in the test tube is ensured to be 5cm higher than the cell block mass at the bottom in the transparence process; wherein, adding a transparent reagent-indicating liquid device to add dimethylbenzene into the test tube;
s7, paraffin soaking: adding liquid paraffin, heating the test tube, stirring, naturally precipitating, repeatedly stirring and precipitating, heating the test tube after the test tube is separated, centrifuging, soaking the paraffin for 1.5 hours, wherein 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 a cell block at the bottom;
wherein, adding the liquid paraffin means adding the liquid paraffin into the test tube by a paraffin extraction device; when liquid paraffin is added into the test tube, the sleeve heating device controls the heating sleeve to ascend until the positioning bulge of the heating sleeve abuts against the positioning block of the test tube wall, the heating sleeve 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, the sleeve heating device controls the heating sleeve to descend to be separated from the test tube;
s8, taking out the test tube: and (3) detaching or cutting the bottom of the wall of the separation test tube, and stripping the metal sheet to obtain the cell wax block.
4. The method according to claim 3, wherein the tube of the liquid extraction device is connected with clear water, and the tube of the liquid extraction device is washed with clear water before the reagent is added into the test tube and before the liquid is extracted from the test tube.
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| KR20130136345A (en) * | 2012-06-04 | 2013-12-12 | 국립암센터 | Composition for aggregating biological sample |
| CN203616181U (en) * | 2013-11-13 | 2014-05-28 | 何鑫 | Combined type centrifuge tube for preparing cytological wax block with trace of exfoliative cells |
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