CN111442962A - Method for manufacturing pathological tissue section - Google Patents
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- CN111442962A CN111442962A CN202010356172.8A CN202010356172A CN111442962A CN 111442962 A CN111442962 A CN 111442962A CN 202010356172 A CN202010356172 A CN 202010356172A CN 111442962 A CN111442962 A CN 111442962A
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- 230000001575 pathological effect Effects 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 47
- 239000012188 paraffin wax Substances 0.000 claims abstract description 28
- 238000005406 washing Methods 0.000 claims abstract description 24
- 239000001993 wax Substances 0.000 claims abstract description 24
- 238000004043 dyeing Methods 0.000 claims abstract description 23
- 230000005540 biological transmission Effects 0.000 claims abstract description 22
- 238000007598 dipping method Methods 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 79
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 28
- 235000019441 ethanol Nutrition 0.000 claims description 28
- 239000008096 xylene Substances 0.000 claims description 18
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 10
- 239000008098 formaldehyde solution Substances 0.000 claims description 10
- 238000002791 soaking Methods 0.000 claims description 10
- 238000004140 cleaning Methods 0.000 claims description 9
- 239000011521 glass Substances 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 6
- 238000010171 animal model Methods 0.000 claims description 5
- 238000007796 conventional method Methods 0.000 claims description 5
- 230000007935 neutral effect Effects 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 5
- 238000007790 scraping Methods 0.000 claims description 3
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 abstract description 6
- 230000018044 dehydration Effects 0.000 abstract description 5
- 238000006297 dehydration reaction Methods 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 3
- 238000001574 biopsy Methods 0.000 abstract description 2
- 238000005520 cutting process Methods 0.000 abstract description 2
- 238000001035 drying Methods 0.000 abstract description 2
- 238000004018 waxing Methods 0.000 abstract description 2
- 239000000853 adhesive Substances 0.000 description 9
- 230000001070 adhesive effect Effects 0.000 description 9
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical group C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 6
- 230000006872 improvement Effects 0.000 description 4
- 230000000249 desinfective effect Effects 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 239000011087 paperboard Substances 0.000 description 3
- 238000009966 trimming Methods 0.000 description 3
- 238000009835 boiling Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005485 electric heating Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
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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
- G01N1/286—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
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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
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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
- G01N1/286—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
- G01N2001/2873—Cutting or cleaving
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Abstract
The invention discloses a method for manufacturing pathological tissue slices, which comprises the following steps: the method comprises the following auxiliary operations of biopsy taking, tissue fixation, tissue washing, dehydration, transparence, wax dipping, embedding, slicing, chip mounting, chip drying, dewaxing, water seepage, water washing, dyeing and the like, and improves the manufacturing quality of the tissue slice and the accuracy of later-stage pathological experiment results. Such as in the steps of waxing and embedding: firstly, the paraffin block is decocted for 2-3h at the temperature of 50-60 ℃, so that the problem of air bubbles after the paraffin is fixed can be greatly avoided, and the yield of tissue slices is improved; through improving the slicer into the mode of automatic transmission tissue slice, cancelled with the step that the instrument of tweezers and writing brush class was transfered the tissue of will cutting into slices, reduced tissue slice's pollution, improved later stage experiment degree of accuracy, make the slicer can cut into slices in succession simultaneously, improve the efficiency of whole preparation.
Description
Technical Field
The invention relates to the technical field of pathology, in particular to a method for manufacturing a pathological tissue section.
Background
The pathological section is one kind of pathological specimen, and is prepared through taking some size animal or pathological tissue, making pathological section in pathological histology, embedding the pathological tissue in paraffin block, slicing in slicer, staining with hematoxylin-eosin (H-E), further examining the pathological change with microscope, and final pathological diagnosis. Pathological sections are pathological tissue structures through observation of cell levels, and the pathological tissue manufacturing process has multiple steps and extremely small section thickness, so that the requirements on the manufacturing quality of tissues and sections are high. The main problems in the current pathological tissue manufacturing process are that after the wax block is embedded, bubbles exist in the wax block, more unqualified slices exist, the quality of tissue slices is influenced, and the rate of finished products of the slices is reduced; on the other hand, after slicing through a conventional slicing machine, an operator transfers sliced tissues by using forceps and tools such as a hairbrush, the sliced tissues are easily polluted by the mode, the later experimental result is influenced, and meanwhile, the slicing machine needs to be continuously started and stopped by the method, so that the slicing efficiency is reduced.
Disclosure of Invention
In view of the above problems, the present invention provides a method for preparing pathological tissue section, comprising the following steps: the method has the advantages that the auxiliary operations of biopsy taking, tissue fixation, tissue washing, dehydration, transparence, wax dipping, embedding, slicing, surface mounting, sheet drying, dewaxing, water seepage, water washing, dyeing and the like are performed, the manufacturing quality of the tissue section and the accuracy of later-stage pathological experiment results are improved, and the pathological section slicing efficiency and the slicing quality are improved through the improvement of the microtome.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a method for manufacturing pathological tissue slices is characterized by comprising the following steps:
a. obtaining a living body: selecting an experimental animal or a diseased tissue after surgical excision, and obtaining an isolated tissue after conventional disinfection;
b. tissue fixation: completely soaking the tissue in 10% neutral formaldehyde solution for 24 h;
c. tissue washing: washing the formaldehyde solution on the fixed tissue for 1-2h by using pure water;
d. and (3) dehydrating: dehydrating the washed tissue in 80% ethanol for 1.5-2h, 95% ethanol for 1-1.5h, and 100% ethanol for 0.5-1 h;
e. and (3) transparency: immersing the dehydrated tissue in dimethylbenzene I for 0.5-1h and dimethylbenzene II for 0.3-0.5h in sequence for transparent treatment;
f. wax dipping and embedding: firstly, decocting a paraffin block at the temperature of 50-60 ℃ for 2-3h, putting the decocted paraffin and the transparent tissue into an electric thermostat at the temperature of 55-60 ℃ for 2-2.5h to soak the paraffin, and embedding to obtain a pathological wax block;
g. slicing: slicing pathological wax blocks by a slicing machine, wherein the slicing thickness is 2-4 um;
the slicing machine comprises a slope cutter seat and an auxiliary device, wherein a blade is arranged at the top of the slope cutter seat, a transmission device is arranged on the slope of the slope cutter seat, the bottom end of the blade extends to the surface of the transmission device, a water bath box is arranged at the bottom of the slope cutter seat, the bottom end of the transmission device extends into the solution of the water bath box, and the sliced pathological tissues are directly immersed into the water bath box at the temperature of 40-45 ℃ through the transmission device;
h. paster and baking sheet
Selecting complete tissue slices in a water bath box, carrying out surface mounting on a glass slide, and placing the cut tissue slices in a thermostat at 60-65 ℃ for 3-5h until the tissue slices are dried;
i. dewaxing
Sequentially dewaxing the dried slices in xylene I and xylene II solutions for 0.2 to 0.3h respectively;
j. water seepage and washing
Soaking the dewaxed slices in absolute ethyl alcohol, 95% ethyl alcohol and 80% ethyl alcohol for 2-3min, washing with pure water, and standing in an anhydrous state;
k. dyeing process
And dyeing the washed slices according to a conventional method, and sealing the slices for later use after dyeing.
Preferably, in step g, the transmission device comprises an upper roller and a lower roller which are arranged on the inclined plane of the inclined plane tool apron, a transmission belt is sleeved on the upper roller and the lower roller, and a micro motor is arranged on one side of the upper roller.
Preferably, a cleaning device is arranged on one side, close to the inclined plane tool apron, of the lower roller wheel and abuts against the surface of the conveying belt.
Preferably, a rubber scraping blade is arranged on the cleaning device and is in surface contact with the conveying belt.
Preferably, the dyeing time in step k is 8-10 min.
The invention has the beneficial effects that: (1) through the complete manufacturing steps of the invention, the manufacturing quality of the tissue section and the accuracy of the later pathological experiment result are improved. (2) In the steps of wax dipping and embedding: firstly, the paraffin block is decocted for 2-3h at the temperature of 50-60 ℃, so that the problem of air bubbles after the paraffin is fixed can be greatly avoided, and the yield of tissue slices is improved. (3) Through the improvement to the slicer, cancelled the step that will cut into slices the tissue with the instrument of tweezers and writing brush class and transmit, reduced tissue slice's pollution, improved the later stage experiment degree of accuracy, make the slicer cut into slices in succession simultaneously, improved the efficiency of whole preparation.
Drawings
Fig. 1 is a schematic perspective view of a microtome according to the present invention.
FIG. 2 is a left side view of the bevel blade holder and the water bath box of the present invention.
FIG. 3 is an enlarged view of FIG. 2A of the present invention.
FIG. 4 is an enlarged view of FIG. 2B of the present invention.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the technical solution of the present invention is further described below with reference to fig. 1 to 4 and examples one to three.
Example one
A method for manufacturing pathological tissue slices comprises the following steps:
a. obtaining a living body, namely selecting an experimental animal or a diseased tissue after surgical excision, conventionally disinfecting to obtain an isolated tissue, and trimming the isolated tissue to a specification of 10 × 10 × 3 mm;
b. tissue fixation: attaching the trimmed isolated tissues to a paperboard, and completely soaking the whole in 10% neutral formaldehyde solution for 24 h;
c. tissue washing: washing the formaldehyde solution on the fixed tissue for 1.5h by using pure water;
d. and (3) dehydrating: dehydrating the washed tissue in 80% ethanol for 1.5h, 95% ethanol for 1h and 100% ethanol for 0.5h in sequence, removing water in the tissue, and because water and paraffin cannot be mutually dissolved, affecting the later-stage embedding quality, the later-stage embedding quality of the tissue can be improved by gradient ethanol dehydration;
e. and (3) transparency: the dehydrated tissue is immersed in dimethylbenzene I for 0.5h and xylene II for 0.3h in sequence for carrying out transparent treatment, and the steps are as follows: because the gradient ethanol for dehydration is not dissolved with the paraffin embedded at the later stage, and the xylene can be dissolved in the ethanol and the paraffin, the transition of the xylene is carried out after the dehydration, when the tissue contains the xylene, the light can penetrate through the tissue, the tissue can present transparent states of different degrees, and the diversity and the transparent quality of the transparency are improved;
f. wax dipping and embedding: firstly, a paraffin block is decocted for 2 hours at the temperature of 50 ℃, bubbles can be eliminated to the maximum extent after tissue slices are fixed by paraffin, the quality of subsequent slices is influenced, the decocted paraffin and transparent tissues are put into an electric heating thermostat at the temperature of 55 ℃ for 2 hours for waxing, then the paraffin and the transparent tissues are immediately put into cold water to be buried to obtain pathological wax blocks, and the pathological wax blocks are put into a refrigerator for refrigerating and storing;
g. slicing: slicing pathological wax blocks by a slicing machine 1, wherein the slicing thickness is 2 um;
the further improvement lies in that: the slicing machine 1 comprises an inclined plane cutter holder 2 and an auxiliary device, wherein a blade 21 is arranged at the top of the inclined plane cutter holder 2, a transmission device 3 is arranged on an inclined plane of the inclined plane cutter holder 2, the bottom end of the blade 21 extends to the surface of the transmission device 3, a water bath box 4 is arranged at the bottom of the inclined plane cutter holder 2, the bottom end of the transmission device 3 extends into a solution of the water bath box 4, and the sliced pathological tissues are directly immersed into the water bath box 4 at 40 ℃ through the transmission device 3;
when slicing, the tissue slices cut by the blade 21 are transmitted to the water bath box 4 through the transmission device 3, so that the step that the tissue slices are transferred by using tweezers and pencils is reduced, the pollution of the tissue slices is reduced, the later-stage experimental accuracy is improved, meanwhile, the slicer 1 can continuously slice, and the integral manufacturing efficiency is improved; the bottom end of the transmission device 3 extends into the solution of the water bath box 4, so that tissue slices are prevented from being adhered to the transmission device 3, the tissue slices are conveniently separated from the transmission device 3, and the continuity of the overall work of the cutter holder is improved;
further to facilitate the transport of the tissue slices: the conveying device 3 comprises an upper roller 31 and a lower roller 32 which are arranged on the inclined surface of the inclined surface tool apron 2, a conveying belt 33 is sleeved on the upper roller 31 and the lower roller 32, a micro motor 34 is arranged on one side of the upper roller 31, when the conveying device works, the micro motor 34 is started to drive the upper roller 31, the conveying belt 33 and the lower roller 32 to rotate, and tissue slices cut by the blade 21 are conveyed into a solution of the water bath box 4 along the conveying belt 33, so that the conveying device is simple and rapid, and the slicing work is stable;
further, in order to clean the water bath solution on the conveying belt 33 conveniently, the cleaning device 5 is arranged on one side, close to the inclined plane tool apron 2, of the lower roller 32, the cleaning device 5 abuts against the surface of the conveying belt 33, when the cleaning device works, the solution attached to the conveying belt 33 in the water bath box 4 is cleaned, the solution is prevented from being thrown out by the conveying belt 33 in the rotating process, the solution is wasted, the working environment is influenced, meanwhile, the tissue slices which are not separated from the conveying belt 33 are cleaned conveniently, and the influence on subsequent tissue slices is eliminated;
further, in order to better clean the water bath solution or the slices on the conveying belt 33, the cleaning device 5 is provided with a rubber scraper 51, the rubber scraper 51 is in contact with the surface of the conveying belt 33, the cutting degree of the rubber scraper 51 and the surface of the conveying belt 33 is better, the solution and the adhered tissue slices on the conveying belt 33 can be thoroughly cleaned, and the use quality and the slicing efficiency of the slicer 1 are improved;
the operation method of other parts of the slicer is the same as the use method of the commercial slicer;
h. paster and baking sheet
Selecting complete tissue slices in the water bath box 4, selecting a clean glass slide, dripping a drop of adhesive in the center of the glass slide, smearing the adhesive to form a thin layer uniformly, then pasting the tissue slices on the adhesive, and putting the tissue slices into a 60 ℃ thermostat for 3 hours after the tissue slices are cut to be dried;
i. dewaxing
In order to facilitate later-stage dyeing, the dried slices are sequentially dewaxed in xylene I and xylene II solutions for 0.2h respectively;
j. water seepage and washing
Soaking the dewaxed slices in absolute ethyl alcohol, 95% ethyl alcohol and 80% ethyl alcohol for 2min, washing with pure water, and standing in an anhydrous state;
k. dyeing process
And dyeing the washed section according to a conventional method, wherein a dyeing agent is preferably hematoxylin, the dyeing time is 8min, and sealing the section for later use after dyeing.
Example two
A method for manufacturing pathological tissue slices comprises the following steps:
a. obtaining a living body, namely selecting an experimental animal or a diseased tissue after surgical excision, conventionally disinfecting to obtain an isolated tissue, and trimming the isolated tissue to 8 × 8-8 × 3mm specification;
b. tissue fixation: attaching the trimmed isolated tissues to a paperboard, and completely soaking the whole in 10% neutral formaldehyde solution for 24 h;
c. tissue washing: washing the formaldehyde solution on the fixed tissue for 1.5h by using pure water;
d. and (3) dehydrating: dehydrating the washed tissue in 80% ethanol for 1.5h, 95% ethanol for 1.2h, and 100% ethanol for 0.7h, removing water from the tissue, and improving the embedding quality of the tissue at later stage;
e. and (3) transparency: sequentially immersing the dehydrated tissue in xylene I for 0.8h and xylene II for 0.5h for carrying out transparent treatment;
f. wax dipping and embedding: firstly, boiling a paraffin block at 55 ℃ for 2h, putting the boiled paraffin and the transparent tissue into a 58 ℃ electric thermostat for 2h to soak the paraffin, then immediately putting the paraffin and the transparent tissue into cold water to be embedded to obtain pathological wax blocks, and putting the pathological wax blocks into a refrigerator for refrigerating and storing;
g. slicing: slicing pathological wax blocks by a slicer 1 (the slicing principle is the same as that in the first embodiment), wherein the slicing thickness is 3 um;
h. paster and baking sheet
Selecting complete tissue slices in the water bath box 4, selecting a clean glass slide, dripping a drop of adhesive in the center of the glass slide, smearing the adhesive to form a thin layer uniformly, then pasting the tissue slices on the adhesive, and placing the tissue slices in a 65 ℃ thermostat for 4 hours until the tissue slices are dried after being cut;
i. dewaxing
Sequentially dewaxing the dried slices in xylene I and xylene II solutions for 0.3h respectively;
j. water seepage and washing
Soaking the dewaxed slices in absolute ethyl alcohol, 95% ethyl alcohol and 80% ethyl alcohol for 3min, washing with pure water, and standing in an anhydrous state;
k. dyeing process
And dyeing the washed section according to a conventional method, wherein a dyeing agent is preferably hematoxylin, the dyeing time is 10min, and sealing the section for later use after dyeing.
EXAMPLE III
A method for manufacturing pathological tissue slices comprises the following steps:
a. obtaining a living body, namely selecting an experimental animal or a diseased tissue after surgical excision, conventionally disinfecting to obtain an isolated tissue, and trimming the isolated tissue to 8 × 8-8 × 3mm specification;
b. tissue fixation: attaching the trimmed isolated tissues to a paperboard, and completely soaking the whole in 10% neutral formaldehyde solution for 24 h;
c. tissue washing: washing the formaldehyde solution on the fixed tissue for 2 hours by using pure water;
d. and (3) dehydrating: dehydrating the washed tissue in 80% ethanol for 2h, 95% ethanol for 1.5h, and 100% ethanol for 1h to remove water in the tissue, thereby improving the quality of embedding in later stage of tissue;
e. and (3) transparency: sequentially immersing the dehydrated tissue in xylene I for 1 hour and xylene II for 0.5 hour for transparent treatment;
f. wax dipping and embedding: firstly, boiling a paraffin block at 55 ℃ for 2.5h, putting the boiled paraffin and the transparent tissue into an electric thermostat at 60 ℃ for 2.5h to soak the paraffin, then immediately putting the paraffin and the transparent tissue into cold water to be embedded to obtain a pathological wax block, and putting the pathological wax block into a refrigerator for cold storage;
g. slicing: slicing pathological wax blocks by a slicer 1 (the slicing principle is the same as that in the first embodiment), wherein the slicing thickness is 4 um;
h. paster and baking sheet
Selecting complete tissue slices in the water bath box 4, selecting a clean glass slide, dripping a drop of adhesive in the center of the glass slide, smearing the adhesive to form a thin layer uniformly, then pasting the tissue slices on the adhesive, and placing the tissue slices in a 65 ℃ thermostat for 3.5 hours until the tissue slices are dried after being cut;
i. dewaxing
Sequentially dewaxing the dried slices in xylene I and xylene II solutions for 0.3h respectively;
j. water seepage and washing
Soaking the dewaxed slices in absolute ethyl alcohol, 95% ethyl alcohol and 80% ethyl alcohol for 3min, washing with pure water, and standing in an anhydrous state;
k. dyeing process
And dyeing the washed section according to a conventional method, wherein a dyeing agent is preferably hematoxylin, the dyeing time is 8min, and sealing the section for later use after dyeing.
According to the specific implementation mode of the embodiment, the method for preparing the pathological tissue section disclosed by the invention can obviously improve the preparation quality of the tissue section and the accuracy of the later pathological experiment result; secondly, in the steps of wax dipping and embedding: firstly, the paraffin block is decocted for 2-3h at the temperature of 50-60 ℃, so that the problem of air bubbles after the paraffin is fixed can be greatly avoided, and the yield of tissue slices is improved; finally, through the improvement of the microtome, the step of transferring the sliced tissues by using forceps and hairbrush tools is eliminated, the pollution of the tissue slices is reduced, the later-stage experimental accuracy is improved, meanwhile, the microtome can continuously slice, and the manufacturing efficiency of the whole tissue slices is improved.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (5)
1. A method for manufacturing pathological tissue slices is characterized by comprising the following steps:
a. obtaining a living body: selecting an experimental animal or a diseased tissue after surgical excision, and obtaining an isolated tissue after conventional disinfection;
b. tissue fixation: completely soaking the tissue in 10% neutral formaldehyde solution for 24 h;
c. tissue washing: washing the formaldehyde solution on the fixed tissue for 1-2h by using pure water;
d. and (3) dehydrating: dehydrating the washed tissue in 80% ethanol for 1.5-2h, 95% ethanol for 1-1.5h, and 100% ethanol for 0.5-1 h;
e. and (3) transparency: immersing the dehydrated tissue in dimethylbenzene I for 0.5-1h and dimethylbenzene II for 0.3-0.5h in sequence for transparent treatment;
f. wax dipping and embedding: firstly, decocting a paraffin block at the temperature of 50-60 ℃ for 2-3h, putting the decocted paraffin and the transparent tissue into an electric thermostat at the temperature of 55-60 ℃ for 2-2.5h to soak the paraffin, and embedding to obtain a pathological wax block;
g. slicing: slicing pathological wax blocks by a slicing machine (1) to obtain slices with the thickness of 2-4 um;
the slicing machine (1) comprises an inclined plane cutter holder (2) and an auxiliary device, wherein a blade (21) is arranged at the top of the inclined plane cutter holder (2), a transmission device (3) is arranged on an inclined plane of the inclined plane cutter holder (2), the bottom end of the blade (21) extends to the surface of the transmission device (3), a water bath box (4) is arranged at the bottom of the inclined plane cutter holder (2), the bottom end of the transmission device (3) extends into a solution of the water bath box (4), and the sliced pathological tissues are directly immersed into the water bath box (4) at the temperature of 40-45 ℃ through the transmission device (3);
h. paster and baking sheet
Selecting complete tissue slices in the water bath box (4), carrying out surface mounting on the glass slide, and placing the cut tissue slices in a thermostat at 60-65 ℃ for 3-5h until the tissue slices are dried;
i. dewaxing
Sequentially dewaxing the dried slices in xylene I and xylene II solutions for 0.2 to 0.3h respectively;
j. water seepage and washing
Soaking the dewaxed slices in absolute ethyl alcohol, 95% ethyl alcohol and 80% ethyl alcohol for 2-3min, washing with pure water, and standing in an anhydrous state;
k. dyeing process
And dyeing the washed slices according to a conventional method, and sealing the slices for later use after dyeing.
2. The method of claim 1, wherein the method comprises: in step g, the transmission device (3) comprises an upper roller (31) and a lower roller (32) which are arranged on the inclined surface of the inclined surface tool apron (2), a transmission belt (33) is sleeved on the upper roller (31) and the lower roller (32), and a micro motor (34) is arranged on one side of the upper roller (31).
3. The method of claim 2, wherein the method comprises: one side of the lower roller (32) close to the inclined plane tool apron (2) is provided with a cleaning device (5), and the cleaning device (5) is abutted to the surface of the transmission belt (33).
4. The method of claim 3, wherein the method comprises: the cleaning device (5) is provided with a rubber scraping blade (51), and the rubber scraping blade (51) is in surface contact with the conveying belt (33).
5. The method of claim 4, wherein the method further comprises: the dyeing time in step k is 8-10 min.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113567482A (en) * | 2021-07-16 | 2021-10-29 | 江南大学 | Method for monitoring internal component structure and distribution of grains in cooking process |
| CN114689407A (en) * | 2022-04-14 | 2022-07-01 | 四川大学华西医院 | Method for manufacturing paraffin section of animal microtissue |
| CN117571363A (en) * | 2024-01-15 | 2024-02-20 | 苏州大学附属第二医院 | Paraffin sample section processing apparatus of pathology department |
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2020
- 2020-04-29 CN CN202010356172.8A patent/CN111442962A/en not_active Withdrawn
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113567482A (en) * | 2021-07-16 | 2021-10-29 | 江南大学 | Method for monitoring internal component structure and distribution of grains in cooking process |
| CN114689407A (en) * | 2022-04-14 | 2022-07-01 | 四川大学华西医院 | Method for manufacturing paraffin section of animal microtissue |
| CN114689407B (en) * | 2022-04-14 | 2023-05-19 | 四川大学华西医院 | Method for manufacturing animal micro tissue paraffin section |
| CN117571363A (en) * | 2024-01-15 | 2024-02-20 | 苏州大学附属第二医院 | Paraffin sample section processing apparatus of pathology department |
| CN117571363B (en) * | 2024-01-15 | 2024-03-22 | 苏州大学附属第二医院 | Paraffin sample section processing apparatus of pathology department |
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Application publication date: 20200724 |