CN108037278B - The preparation method of immunohistochemistry detection slice - Google Patents
The preparation method of immunohistochemistry detection slice Download PDFInfo
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- CN108037278B CN108037278B CN201711330986.9A CN201711330986A CN108037278B CN 108037278 B CN108037278 B CN 108037278B CN 201711330986 A CN201711330986 A CN 201711330986A CN 108037278 B CN108037278 B CN 108037278B
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Abstract
The invention discloses a kind of preparation methods of immunohistochemistry detection slice, include the following steps：The paraffin section of tissue sample to be detected is obtained, and the paraffin section is laid on plastic foil, paraffin section forms the paraffin section for attaching plastic foil with plastic foil tight bond, bakes piece；Paraffin section punching to the attaching plastic foil after the roasting piece, obtains the paraffin section of the circular attaching plastic foil of at least a piece of a diameter of 1~30mm；The circular paraffin section for attaching plastic foil is transferred in ELISA Plate, it is a piece of per hole, the circular hydrophilic tissue slice for attaching plastic foil is obtained after the aquation that dewaxes；The circular hydrophilic tissue slice for attaching plastic foil is dyed, you can.The present invention by inventor creatively by plastic foil together with paraffin section tight bond, obtain multiple paraffin sections using punching, final realize prepares multiple slices using the paraffin section of a piece of tissue to be detected, while meeting the needs of multinomial detection.
Description
Technical field
The present invention relates to biotechnologies, more particularly to a kind of preparation method of immunohistochemistry detection slice.
Background technology
One kind that cut sections for microscopic examination (pathological examination) refer to pathologic finding, is used to check body device
The Pathomorphology method of pathological change in official, tissue or cell.Currently, cut sections for microscopic examination have been widely used in clinical work
Make, scientific research, the preparation of microscopy slice plays key effect for the diagnosis of result.The preparation of traditional microscopy slice is mainly
It takes the pathologic tissue packing that size is 2.0cm × 2.0cm × 0.3cm in paraffin, slice acquisition paraffin is then carried out to it and is cut
Piece, then the paraffin section of the full wafer of gained is attached on glass slide and is dyed, the paraffin section per whole piece is only capable of carrying out
Primary dyeing prepares a microscopy slice, finally obtains a result.
Since in actually detected, clinical tissue specimen samples are few, in addition the complexity of microsection manufacture, and need to same
Tissue sample to be detected carries out a variety of antigens or antibody or other relevant detections, needs to use multiple detection slices, therefore,
Traditional detection slice preparation method is difficult to meet the needs of follow-up diversity detection.
Invention content
Based on this, it is necessary to which multiple inspections can be obtained by providing a kind of paraffin section by a piece of tissue sample to be detected
The method for surveying slice.
A kind of preparation method of immunohistochemistry detection slice, includes the following steps：
The paraffin section of tissue sample to be detected is obtained, and the paraffin section is laid on plastic foil, paraffin section
The paraffin section for attaching plastic foil is formed with plastic foil tight bond, bakes piece；
The paraffin section punching that plastic foil is attached to gained, obtains the circular attaching of at least one a diameter of 1~30mm
The paraffin section of plastic foil；
The circular paraffin section for attaching plastic foil is transferred in ELISA Plate, it is a piece of per hole, after the aquation that dewaxes
Obtain the circular hydrophilic tissue slice for attaching plastic foil；
The circular hydrophilic tissue slice for attaching plastic foil is dyed, you can.
In wherein some embodiments, a diameter of 3~20mm of the circular paraffin section for attaching plastic foil.
In wherein some embodiments, the plastic foil is the plastic foil that tolerable temperature is not less than 265 DEG C；The plastic foil
Brittleness temperature be not higher than -196 DEG C of plastic foil.
In wherein some embodiments, the plastic foil is polyimide plastic film or poly- naphthalene ester plastic foil (2,6 naphthalene diformazans
Sour glycol ester film).
In wherein some embodiments, the thickness of the plastic foil is 6~250 μm.
In wherein some embodiments, the thickness of the plastic foil is 30~250 μm.
In wherein some embodiments, the condition of the roasting piece is：Temperature is 60~65 DEG C, the time is 25~30mi n.
In wherein some embodiments, the plastic foil is also pre-processed as follows：First applied on the surface of the plastic foil
It is 5 to cover volume ratio：It is dry after the egg white of (1~3), the mixture of anionic polyelectrolyte aqueous solution then pre- in the plastic foil
The surface for being laid with paraffin section coats drying after cationic polyelectrolyte solution.
In wherein some embodiments, the anionic polymer can be polyacrylic acid, polymethylacrylic acid, polystyrene
Sulfonic acid, polyvinyl sulfonic acid or polyvinyl；The cationic polyelectrolyte can be polyvinylamine, polyvinyl pyridine, polydiene
Third alkyl dimethyl ammonium chloride or polyethyleneimine.
In wherein some embodiments, the plastic foil is equal after coating the mixture, cationic polyelectrolyte solution
It need to be transferred in sealing container, be respectively less than in boiling point, surface tension in the steam ambient of the solvent of water and carry out steam treatment.
Adaptive immune groupization detection slice, which can be placed directly in microplate reader, after dyeing of the embodiment of the present invention quantitatively detects, or
Person from be transferred on glass slide in ELISA Plate, mounting is placed under microscope and carries out qualitative detection.
Compared with prior art, the invention has the advantages that：
The present invention by inventor creatively by plastic foil together with paraffin section tight bond, obtained using punching
Multiple paraffin sections are obtained, in particular, a diameter of 3~20mm by controlling the circular paraffin section for attaching plastic foil, it can
It realizes that the circular paraffin section for attaching plastic foil of multi-disc carries out different dyeing processing in ELISA Plate, makes itself and ELISA Plate
Aperture matches, and final realize prepares multiple slices using the paraffin section of a piece of tissue to be detected, while meeting multinomial detection
Demand.The present invention dexterously combines ELISA Plate, and dyeing processing is carried out to the small dimension paraffin section of acquisition, reduces reagent and uses
Amount is also easy to control humidity, avoids reagent evaporation, tissue dry to cause harmful effect to coloration result, quickly obtains height
The microscopy of quality is sliced.
The plastic foil of the application is preferably the plastic foil that tolerable temperature is not less than 265 DEG C, and plastics can be avoided using the film
The combination of film and histotomy causes histotomy to tear in subsequent heat processing procedure because of the thermal expansion of plastic foil,
Will not lead to that histotomy is caused to occur folding to influence follow-up coloring and observation effect because of the cooling meat of plastic foil
Fruit.
The plastic foil of the application is preferably the plastic foil that brittleness temperature is not higher than -196 DEG C, can avoid punching using the film
In the process stress and there is plastic foil cracking, the white phenomenon of fine fisssure and top, realize that plastic foil is consistent with histotomy stress, shape
Become convergent, to ensure that the stability of plastic foil and histotomy combination, the homogeneity of thickness, finally ensures chromatin
Amount.
Description of the drawings
Fig. 1 is that histotomy with card punch is broken into small pieces and be placed in ELISA Plate and carries out immunochemistry by the embodiment of the present invention
The schematic diagram of colouring method；
Fig. 2 is the result signal that using implementation of the present invention breast tumor tissue sections are carried out with immune group chemical staining
Figure.
Specific implementation mode
The preparation method of the immunohistochemistry detection slice of the present invention is made below in conjunction with specific embodiment further details of
Explanation.In order to enable the preparation method of immunohistochemistry detection slice provided in an embodiment of the present invention is clearer, now with to mammary gland
Tissue sample carries out film-making, determines that breast cancer molecular subtypes are illustrated after microscopy, but it should be noted that institute of the present invention
Scheme to be protected is not limited to following each examples.It is understood that the paraffin section described in the embodiment of the present invention is using conventional
The paraffin section that method (such as including materials, fixation, washing and dehydration, transparent, waxdip and embedding, slice) is prepared is thick
Degree is 3~5 μm.
In traditional sample of breast tissue detection, generally requires and carry out breast cancer Pathologic specimen respectively using six kinds of markers
Carry out immunohistochemical staining, including estrogen receptor (ER), progesterone receptor (PR), HER2, CK5/6, epidermal growth factor receptor
Body (EGFR) and Ki-67.It is divided into epithelium type (being divided to A, B two kinds) according to the expression of this six kinds of markers, HER2 crosses table
Up to type and substrate template, the selection and patient's prognosis of division and the clinical application of different subtype are closely related, have important
Clinical reference value.If according to traditional flaking method, six kinds of markers are detected simultaneously and at least need six conventional rule
The paraffin section of lattice, is respectively placed on glass slide, and immunochemistry dyeing is carried out with different antibodies, final to prepare six observation pieces,
The paraffin section of i.e. each conventional specification is simply possible to use in a kind of marker of detection, can thus increase to the demand of pathological tissue
Greatly, relevant treatment reagent dosage increases.
And use method provided in an embodiment of the present invention, it is only necessary to six kinds of marks can be completed using a piece of paraffin tissue sections
The detection of object.Commercially available card punch may be used in device used by punching of the embodiment of the present invention.It is used in the embodiment of the present invention
Control, primary antibody, fluorescence secondary antibody are conventional class, wherein positive reference substance include ER positive reference substances, PR positive reference substances,
HER2 positive reference substances, CK5/6 positive reference substances, EGFR positive reference substances, Ki-67 positive reference substances；Primary antibody includes anti-ER anti-
Body, anti-PR antibody, Anti-HER 2, anti-CK5/6 antibody, anti-egfr antibodies, anti-Ki-67 antibody；Fluorescence secondary antibody includes anti-ER anti-
The fluorescence secondary antibody, anti-of the fluorescence secondary antibody of body, the fluorescence secondary antibody of anti-PR antibody, the fluorescence secondary antibody of Anti-HER 2, anti-CK5/6 antibody
The fluorescence secondary antibody of the fluorescence secondary antibody of EGFR antibody, anti-Ki-67 antibody.It is understood that it is also ready for antibody dilute solution, institute
Some primary antibodies are held using the Brown Glass Brown glass bottles and jars only of black caps, and fluorescence secondary antibody is held with the Brown Glass Brown glass bottles and jars only of red cap, Antibody Concentration
The black PE plastic bottles of solution black caps are held, and antibody diluent is held with the white PE plastic bottles of red cap.
Embodiment 1
The present embodiment provides a kind of preparation methods of immunohistochemistry detection slice, include the following steps：
Step 1 obtains the paraffin section of sample of breast tissue to be detected, and the paraffin section is laid in polyamides Asia
On amine plastic foil (commercially available, tolerable temperature is 400 DEG C, subzero 200 DEG C of brittle temperature), paraffin section and plastic foil tight bond shape
At the paraffin section for attaching plastic foil, in 60 DEG C of roasting piece 30min after flattening-out；The thickness for the polyimide plastic film that this step uses
It is 30 μm；Flattening-out mode described in this step is that in breast cancer paraffin section, roller is dynamic back and forth using rubber cylinder；
Step 2, the paraffin section that plastic foil is pasted with to gained punch, and obtaining 18, (total 6 processing, are each handled
It is repeated 3 times) the circular paraffin section for being pasted with plastic foil of a diameter of 3mm；
The paraffin section fritter for being pasted with plastic foil is transferred in ELISA Plate hole by step 3, a piece of per hole, through de-
The circular hydrophilic tissue slice for attaching plastic foil is obtained after wax aquation；In order to improve the accuracy of detection, which also sets
There are ER positive reference substances, PR positive reference substances, HER2 positive reference substances, CK5/6 positive controls, EGFR positive controls, Ki-67 sun
Property control, each control is repeated once；The processing of each of ELISA Plate is marked during subsequent immunostaining, enzyme mark
Removing the work office can refer to as follows everywhere on plate：
The circular hydrophilic tissue slice for attaching plastic foil obtained by step 3 is carried out immune group chemistry dye by step 4
Color, and transferred them on glass slide after the dyeing, it can mounting without removing the round plastic film attached.
Immunochemistry dyeing described in this step includes antigen retrieval, site closing plus primary antibody plus two contragradience of fluorescence
Suddenly；
The dewaxing aquation includes：The paraffin in slice is taken off with dimethylbenzene and graded ethanol and makes histocyte again
It is integrated with water；Wherein, the continuous transparent processing of dimethylbenzene dewaxing 2 times, each 10min；The graded ethanol elution is specifically first
5min is eluted with 100% concentration alcohol, then twice with the elution of 95% concentration alcohol, each 2min；" % " tool described in the step
Body refers to volume ratio；It is understood that after dewaxing aquation, deionized water also can be used and rinse one time；
The antigen retrieval includes：It is added in citrate buffer (pH6.0), makes in each hole into ELISA Plate
With microwave stove heat, first height fire 5min, then in low fire 10min, finally naturally cool to room temperature；It is understood that antigen
It after repairing also needs that PBS buffer solution is added into each hole of ELISA Plate, histotomy be washed, the number generally washed
It is 3 times, each 3min；
It closes in the site：3%H is added into each hole of ELISA Plate2O2(% is percent by volume, and solvent is
PBS buffer solution), it is protected from light and is incubated 10min, deionized water washing；It can be understood that after the closing of site, the tissue in each hole
Slice need to be washed using PBS, and the number of washing is 3 times, each 3min；
Described plus primary antibody, including：Be added dropwise respectively according to label antiestrogenic antibody, antiprogestin antibody and anti-HER2,
The specific antibody of CK5/6, EGFR and Ki-67 antigen (can also be prepared) sealing cover ELISA Plate with sealing plate film by heat-resisting material,
4 DEG C overnight；
Described plus fluorescence secondary antibody, including：ELISA Plate is taken out from refrigerator, is put into PBS and washes 3 times, 5 minutes every time,
After blotting the PBS around tissue, respective fluorescence secondary antibody is added dropwise, is subsequently placed in 37 DEG C of incubators 0.5 hour；
Fluorescence secondary antibody after treatment, takes out ELISA Plate from incubator, is put into PBS buffer solution and continuously washs 3 times, every time
The circular hydrophilic tissue slice overturning for being pasted with plastic foil after the PBS buffer solution for blotting surrounding, is placed in load glass by 5min
On piece makes hydrophilic tissue slice be attached on glass slide, is observed under the microscope in fluorescence microscopy after mounting.
The present embodiment may be used sample of breast tissue of 24 orifice plate pair according to above-mentioned setting and carry out immunochemistry dyeing,
Immunochemistry dyeing, specific schematic diagram can also be carried out to four sample of breast tissue simultaneously using 96 orifice plates with reference to above-mentioned setting
See Fig. 1, wherein 1 is sealing plate film, 2 be the hole of ELISA Plate, and 3 be the paraffin section fritter with plastic foil, and 4 be ELISA Plate (96 holes
Plate).
Sample of breast tissue paraffin section is first laid on plastic foil by the present embodiment, is punched to it after flattening-out, then
Several histotomy small pieces of acquisition are respectively placed in progress immunochemistry dyeing in the hole of ELISA Plate, only with a piece of paraffin
The detection of Multiple Antibodies can be carried out under conditions of slice, it is easy to operate, efficient, cost-effective, and sample of breast tissue stone
Wax, which is sliced to be firmly combined with plastic foil, does not occur obscission.
The plastic foil that the present embodiment is selected is polyimide plastic film, and the tolerable temperature of the plastic foil is more than 265 DEG C, and heat is steady
It is qualitative consistent with tissue sample, the combination of plastic foil and histotomy can be avoided (such as roasting in subsequent heat processing step
Piece, antigen retrieval) cause histotomy to tear because of the thermal expansion of plastic foil in the process, the cooling because of plastic foil will not be caused
It shrinks and histotomy is caused to occur folding to influence follow-up coloring and observing effect.The plastic foil that this example is selected is crisp
Warm-natured plastic foil of the degree not higher than -196 DEG C, can avoid stress in drill process using the film and plastic foil occur and open
Split, the white phenomenon of fine fisssure and top, realize that plastic foil is consistent with histotomy stress, deformation is convergent, to ensure that plastic foil and
The stability of histotomy combination, the homogeneity of thickness, finally ensure dyeing quality.
During the preparation method of the histotomy provided according to the present example prepares 100 histotomies, only 5 tissues
It is sliced and occurs the situation that plastic foil falls off in Liquid-treatment processes；Although this example carries out film-making using small dimension paraffin section,
But sample of breast tissue processing to be detected is uniformly abundant, coloring is good, and microscopic observation result is shown in Fig. 2.
In Fig. 2, Fig. 2A is estrogen receptor (ER) positive control, and Fig. 2 B are that detection sample estrogen receptor (ER) is cloudy
Property, Fig. 2 C are progesterone receptor (PR) color positive control, and it is negative that Fig. 2 D, which are detection sample progesterone receptor (PR), and Fig. 2 E are
HER2 fluorescent staining positive controls, Fig. 2 F are that detection sample HER2 is negative, and Fig. 2 G are that detection sample EGFR is positive, and Fig. 2 H are
It is the positive to detect sample CK5/6, and Fig. 2 I are that Ki-67 fluorescent stainings are positive.The above results show that tissue sample to be detected is base
Ground template breast cancer.
Embodiment 2
The present embodiment is the change case of embodiment 1, and compared with Example 1, variation place is only that：
In step 1, plastic foil selects poly- naphthalene ester plastic foil, and (commercially available, tolerable temperature is 265 DEG C, and brittleness temperature is less than subzero
196 DEG C), it is 65 DEG C to bake piece temperature, and the thickness of polyester plastics film is 250 μm；Flattening-out mode described in this step is using glass
Roller is dynamic back and forth in breast cancer paraffin section for stick；
In step 2, the paraffin section that plastic foil is pasted with to gained punches, and obtains the circular attaching of a diameter of 20mm
There is the paraffin section of plastic foil.
The present embodiment carries out immunofluorescence dyeing using four six orifice plates.
During the preparation method of the histotomy provided according to the present example prepares 100 histotomies, only 5 tissues
It is sliced and occurs the situation that plastic foil falls off in Liquid-treatment processes；Although this example carries out film-making using small dimension paraffin section,
But sample of breast tissue processing to be detected is uniformly abundant, coloring is good, and microscopic observation result is consistent with embodiment 1.
Embodiment 3
The present embodiment is the improvement of embodiment 1, and compared with Example 1, improvements are only that：
In step 1, the plastic foil is also pre-processed as follows：First on the surface of the plastic foil, coating volume ratio is
5：It is dry after 2 egg white, the mixture of anionic polyelectrolyte aqueous solution (mass concentration of the aqueous solution is 0.5%), then
The plastic foil is laid with the surface coating cationic polyelectrolyte solution of paraffin section in advance, and (mass concentration of the aqueous solution is
0.5%) it is dried after, wherein anionic polyelectrolyte is polyacrylic acid, cationic polyelectrolyte is polyvinylamine.Described in this example
Dry is to dry.
This example carries out above-mentioned pretreatment to plastic foil, can not only promote the secured journey that histotomy is combined with plastic foil
Degree, and in processing procedure, plastic foil has good hydrophilicity, and treatment liquid is added into ELISA Plate, (such as PBS is slow
Fliud flushing) when, plastic foil-paraffin section can be good at infiltration in treatment liquid, realize fine treatment effect.Specifically, originally
Inventive embodiments use volume ratio for 5：The egg white of (1~3), the design of mixture of anionic polyelectrolyte aqueous solution, even if
Long time is infiltrated in liquid, which is still stable in the presence of the surface of plastic foil, and this layer of mixture is in plastics
The film layer good water permeability that film surface is formed, will not hinder the treatment effect of histotomy, and further, cation is poly- after re-coating
After electrolyte solution, cationic polyelectrolyte can be by electrostatic interaction and anionic polyelectrolyte physical absorption, while also can
Enough by the protein binding on charge effect and histotomy, to the more secured binding force of histotomy and plastic foil.
Importantly, inventor find, with pretreatment plastic foil carry histotomy also avoid during subsequent operation
Histotomy edge is easy the withered phenomenon of evaporation.
During the preparation method of the histotomy provided according to the present example prepares 100 histotomies, only 3 tissues
It is sliced and occurs the situation that plastic foil falls off in Liquid-treatment processes；Although this example carries out film-making using small dimension paraffin section,
But sample of breast tissue processing to be detected is uniformly abundant, coloring is good, and microscopic observation result is consistent with embodiment 1.
Embodiment 4
The present embodiment is the improvement of embodiment 3, and compared with Example 3, improvements are only that：In step 1, plastics
Film is transferred in sealing container after coating the mixture, cationic polyelectrolyte solution, small in boiling point, surface tension
Steam treatment is carried out in the steam ambient of the solvent (chloroform) of water.
In preprocessing process, if convection drying is (using directly drying or direct low temperature without steam treatment
Drying), then the film layer formed in plastic film after pretreatment can be uneven, this uneven state is to histotomy and modeling
The combined with firmness of material film bring hidden danger, it is easy to the histotomy in subsequent processes be caused to fall off.By for a long time
Grope, inventor infer, this inhomogeneities may be due to caused by " coffee cup " effect.For this purpose, inventor is to coating
Plastic foil afterwards carries out steam treatment, and steam is below the solvent of water from surface tension, boiling point, which can overcome
" coffee cup " effect so that the substance coated on frosting uniformly deposits stratification, finally obtains smooth surface, increase group
The stability of slice and plastic foil is knitted, also, inventor is also found surprisingly that, the hydrophily of plastic foil plays aobvious after steam treatment
The promotion of work can obtain preferable treatment effect in a relatively short period of time in subsequent processing.
During the preparation method of the histotomy provided according to the present example prepares 100 histotomies, do not occur plastics
The situation that film falls off；Although this example carries out film-making using small dimension paraffin section, sample of breast tissue processing to be detected is uniform
Fully, coloring is good, and microscopic observation result is consistent with embodiment 1.
Comparative example 1
This example is the comparative example of embodiment 1, and compared with Example 1, comparison place is only that：The plastic foil used is commercially available
Polycarbonate plastic film, 150 DEG C of tolerable temperature, brittleness temperature are subzero 100 DEG C.
As a result, implementation steps are found for the moment, paraffin section meeting during flattening-out bakes piece of sample of breast tissue to be detected
There is slice with plastics UF membrane to which there are the states of minute bubbles for presentation, and plastic foil deformation is larger in roasting piece, to
So that part tearing state occurs in the paraffin section of docile thereon, the utilization rate of paraffin section is reduced；In addition, in the mistake of punching
Fine fisssure is susceptible in journey, be attached to fine fisssure position paraffin section fritter be susceptible in subsequent liquid processing procedure it is de-
Fall, the histotomy to fall off is easy to happen folding since thickness is smaller, to be unfavorable for subsequently dyeing, be also unfavorable for by its from
It takes out to be transferred on glass slide in ELISA Plate and observe.
Although this example carries out film-making using small dimension paraffin section, sample of breast tissue to be detected processing uniformly fully,
Coloring is good, and microscopic observation result is consistent with embodiment 1, but prepared by the preparation method of the histotomy provided according to the present example
During 100 histotomies, there are 15 histotomies to occur the situation that plastic foil falls off in Liquid-treatment processes.
Comparative example 2
This example is the comparative example of embodiment 1, and compared with Example 1, comparison place is only that：
In step 1, the thickness of polyimide plastic film is 6 μm；
In step 2, a diameter of 1mm of the circular paraffin section for being pasted with plastic foil.
Although sample of breast tissue processing to be detected is uniformly abundant, coloring is good, electric microscopic observation result and embodiment 1
Unanimously, during the preparation method of the histotomy but provided according to the present example prepares 100 histotomies, 10 tissues cut
There is the situation that plastic foil falls off in Liquid-treatment processes in piece.
Comparative example 3
This example is the comparative example of embodiment 1, and comparison place is that step 1 is specifically：Obtain sample of breast tissue to be detected
Paraffin section, and the paraffin section is laid on transparent glassine paper, in 60 DEG C of roasting piece 30min after flattening-out.
As a result, it is easy to appear alice (i.e. sides at the edge of the paraffin section fritter for the attached glassine paper that step 2 punching obtains
There is paraffin section and is detached with glassine paper in edge), to which histotomy easily falls off from glassine paper in subsequent processing, and bake
Paraffin after piece on paraffin section is easily impregnated into glassine paper so that glassine paper hydrophobicity increases, and significantly increases follow-up de-
The duration of wax aquation reduces histotomy treatment effeciency.The preparation method of the histotomy provided according to the present example prepares 100 groups
During knitting slice, there are 90 histotomies to occur the situation that plastic foil falls off in Liquid-treatment processes；In order to realize compared with
Good dewaxing aquation needs the first dimethylbenzene continuous transparent processing of dewaxing 4 times (each 15mi n), once again ethanol elution, specifically first
15mi n are eluted with 100% concentration alcohol, then are eluted 4 times with 95% concentration alcohol, each 5mi n.
Comparative example 4
This example is the comparative example of embodiment 1, and comparison place is that this example is one and first punches the technical solution for baking piece afterwards, right
Than the step of include step 1 and step 2, it is specific as follows：
Step 1 obtains the paraffin section of sample of breast tissue to be detected, and the paraffin section is laid in polyamides Asia
On amine plastic foil, it is punched after flattening-out, to obtain the circular paraffin section for being pasted with plastic foil；
The circular paraffin section for being pasted with plastic foil is placed in 60 DEG C of roasting piece 30mi n by step 2.
The fussy degree that piece not only increases operation is baked again as a result, it has been found that first punching, it is often more important that directly to paraffin section
It carries out punching and easilys lead to the fixed tissue of paraffin occurring marginal laceration, broken with the crushing of paraffin, be unfavorable for its modeling
Expect the tight bond of film.During the preparation method of the histotomy provided according to the present example prepares 100 histotomies, there is 80
It opens histotomy and occurs the situation that plastic foil falls off in Liquid-treatment processes.
Each technical characteristic of embodiment described above can be combined arbitrarily, to keep description succinct, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, it is all considered to be the range of this specification record.
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. a kind of preparation method of immunohistochemistry detection slice, which is characterized in that include the following steps：
The paraffin section of tissue sample to be detected is obtained, and the paraffin section is laid on plastic foil, paraffin section and modeling
Expect that film tight bond forms the paraffin section for attaching plastic foil, bakes piece；
Paraffin section punching to the attaching plastic foil after the roasting piece, obtains the circular of at least a piece of a diameter of 1~30mm
Attach the paraffin section of plastic foil；
The circular paraffin section for attaching plastic foil is transferred in ELISA Plate, it is a piece of per hole, it is obtained after the aquation that dewaxes
The circular hydrophilic tissue slice for attaching plastic foil；
The circular hydrophilic tissue slice for attaching plastic foil is dyed, you can；
The plastic foil is the plastic foil that tolerable temperature is not less than 265 DEG C.
2. the preparation method of immunohistochemistry detection slice according to claim 1, which is characterized in that the circular attaching
A diameter of 3~20mm of the paraffin section of plastic foil.
3. the preparation method of immunohistochemistry according to claim 1 detection slice, which is characterized in that the plastic foil it is crisp
Warm-natured plastic foil of the degree not higher than -196 DEG C.
4. the preparation method of immunohistochemistry detection slice according to claim 3, which is characterized in that the plastic foil is poly-
Acid imide plastic foil or poly- naphthalene ester plastic foil.
5. the preparation method of immunohistochemistry detection slice according to any one of claims 1 to 4, which is characterized in that described
The thickness of plastic foil is 6~250 μm.
6. the preparation method of immunohistochemistry detection slice according to claim 5, which is characterized in that the thickness of the plastic foil
Degree is 30~250 μm.
7. the preparation method of immunohistochemistry detection slice according to any one of claims 1 to 4, which is characterized in that described
The condition of roasting piece is：Temperature is 60~65 DEG C, the time is 25~35min.
8. the preparation method of immunohistochemistry detection slice according to any one of claims 1 to 4, which is characterized in that it is special
Sign is that the plastic foil is also pre-processed as follows：First on the surface of the plastic foil, coating volume ratio is 5：(1~3)
Egg white, anionic polyelectrolyte aqueous solution mixture after it is dry, then be laid in the plastic foil surface of paraffin section in advance and apply
It is dried after covering cationic polyelectrolyte solution.
9. the preparation method of immunohistochemistry detection slice according to claim 8, which is characterized in that the anionic polymerisation
Object can be polyacrylic acid, polymethylacrylic acid, polystyrolsulfon acid, polyvinyl sulfonic acid or polyvinyl.
10. the preparation method of immunohistochemistry detection slice according to claim 8, which is characterized in that the cation is poly-
Electrolyte can be polyvinylamine, polyvinyl pyridine, the third alkyl dimethyl ammonium chloride of polydiene or polyethyleneimine；The plastic foil is applying
It is both needed to be transferred in sealing container after covering the mixture, cationic polyelectrolyte solution, water is respectively less than in boiling point, surface tension
Solvent steam ambient in carry out steam treatment.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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