CN112816295A - Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section - Google Patents
Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section Download PDFInfo
- Publication number
- CN112816295A CN112816295A CN202011614151.8A CN202011614151A CN112816295A CN 112816295 A CN112816295 A CN 112816295A CN 202011614151 A CN202011614151 A CN 202011614151A CN 112816295 A CN112816295 A CN 112816295A
- Authority
- CN
- China
- Prior art keywords
- bone marrow
- marrow tissue
- decalcification
- decalcifying
- tissue
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000001519 tissue Anatomy 0.000 title claims abstract description 70
- 210000001185 bone marrow Anatomy 0.000 title claims abstract description 60
- 238000000034 method Methods 0.000 title claims abstract description 38
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 31
- 239000007788 liquid Substances 0.000 title claims abstract description 23
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 38
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 238000011532 immunohistochemical staining Methods 0.000 claims abstract description 11
- 239000012153 distilled water Substances 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 33
- 238000002791 soaking Methods 0.000 claims description 12
- 230000018044 dehydration Effects 0.000 claims description 9
- 238000006297 dehydration reaction Methods 0.000 claims description 9
- 239000011547 Bouin solution Substances 0.000 claims description 5
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 4
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 claims description 4
- 239000008096 xylene Substances 0.000 claims description 4
- 230000000694 effects Effects 0.000 abstract description 13
- 238000003745 diagnosis Methods 0.000 abstract description 8
- 238000010186 staining Methods 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 235000019441 ethanol Nutrition 0.000 description 20
- 238000005406 washing Methods 0.000 description 20
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 14
- 238000007654 immersion Methods 0.000 description 7
- 230000002055 immunohistochemical effect Effects 0.000 description 7
- 239000002253 acid Substances 0.000 description 6
- 238000004043 dyeing Methods 0.000 description 6
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 230000001575 pathological effect Effects 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 238000000861 blow drying Methods 0.000 description 2
- 230000009920 chelation Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 2
- -1 primary antibody Substances 0.000 description 2
- 239000012744 reinforcing agent Substances 0.000 description 2
- 101100381525 Mus musculus Bcl6 gene Proteins 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000012024 dehydrating agents Substances 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000004821 effect on bone Effects 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Images
Classifications
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a bone marrow tissue decalcifying liquid and a method for preparing a bone marrow tissue paraffin section, wherein the bone marrow tissue decalcifying liquid consists of hydrochloric acid, formaldehyde and distilled water, wherein the hydrochloric acid accounts for 1-3% of the volume percentage concentration of the bone marrow tissue decalcifying liquid, and the formaldehyde accounts for 9-11% of the volume percentage concentration of the bone marrow tissue decalcifying liquid. The invention provides a bone marrow tissue decalcifying liquid and a method for manufacturing a bone marrow tissue paraffin section, the bone marrow tissue specimen is decalcified by adopting the decalcifying liquid, the specimen pretreatment process is shortened, the report time is short, the more timely diagnosis is facilitated, and the obtained HE section shows that the cell morphology is well preserved and the tissue structure is clear; the immunohistochemical staining section shows that the staining effect is excellent, the positioning is accurate, and the background is clean; and the sample does not have the phenomenon of excessive decalcification.
Description
Technical Field
The invention relates to the technical field of pathological detection, in particular to a bone marrow tissue decalcifying liquid and a method for preparing a bone marrow tissue paraffin section.
Background
Bone marrow biopsy is one of the important components in pathological examination, and the diagnosis report thereof is an important basis for disease diagnosis and treatment in clinical departments such as hematology department and the like. At present, two methods, namely a plastic method and a paraffin method, are mainly used for embedding the bone marrow biopsy specimen, wherein the plastic method is not popularized yet because the plastic method has various treatment procedures, the antigen activity is easy to lose, and the immunohistochemical staining is quite difficult, and the paraffin method is mainly used.
The most important link for preparing high-quality bone marrow slices by adopting the paraffin method is the fixation and decalcification of specimen tissues. If the bone marrow specimen tissue is not sufficiently fixed, the morphological structure of the specimen tissue and various components in cells can be damaged in the subsequent link; if the decalcification of the specimen tissue is incomplete, a complete paraffin section cannot be cut, and the section is easy to fall off in the dyeing process, so that the tissue structure is easy to be incomplete artificially.
The decalcification methods commonly used at present include EDTA chelation decalcification, electrolytic decalcification and acid decalcification. However, the EDTA chelation decalcification and the organic weak acid decalcification take a long time to affect the timeliness of treatment; the electrolysis decalcification requires special equipment, and is high in cost and inconvenient; inorganic strong acid decalcification generally has high decalcification speed, is difficult to control in time, needs continuous needling observation, is easy to decalcification excessive, can cause loss of a plurality of antigens and has obvious influence on immunohistochemical staining; the mixed decalcification of strong inorganic acid and organic acid requires treatment of residual acid.
That is, how to fix and decalcify the specimen in a short time, and the prepared section does not affect the staining, and can ensure that the tissue antigen is not lost, and the immunohistochemical staining effect is good, thereby being helpful for the under-lens diagnosis of the pathological tissue, which is always a great problem in the field.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides the bone marrow tissue decalcification solution and the method for preparing the bone marrow tissue paraffin section, which realize the short decalcification time, convenient and timely diagnosis and the good HE dyeing and immunohistochemical dyeing effects.
The invention adopts the following technical scheme:
the invention provides a bone marrow tissue decalcification solution, which consists of hydrochloric acid, formaldehyde and distilled water, wherein the hydrochloric acid accounts for 1-3% of the bone marrow tissue decalcification solution by volume percentage, and the formaldehyde accounts for 9-11% of the bone marrow tissue decalcification solution by volume percentage.
According to research, the decalcification liquid can be used for decalcification of bone marrow tissue specimens quickly, so that the diagnosis is facilitated in time, excessive decalcification and antigen loss are avoided, and the section with good HE (high-grade hematoxylin) dyeing and immunohistochemical dyeing effects is obtained.
Preferably, the hydrochloric acid accounts for 2% of the volume percentage of the bone marrow tissue decalcifying solution, and the formaldehyde accounts for 10% of the volume percentage of the bone marrow tissue decalcifying solution.
In the technical scheme, the hydrochloric acid is diluted hydrochloric acid with the mass fraction of 2-3%.
The invention also provides a method for preparing the paraffin section of the bone marrow tissue, which comprises the step of decalcifying the specimen by adopting the bone marrow tissue decalcifying liquid.
The bone marrow tissue specimen is decalcified by adopting the decalcifying liquid, the pretreatment process of the specimen is shortened, the report time is short, the more timely diagnosis is facilitated, and the obtained HE section shows that the cell morphology is well preserved and the tissue structure is clear; the immunohistochemical staining section shows that the staining effect is excellent, the positioning is accurate, and the background is clean; and the sample does not have the phenomenon of excessive decalcification.
Further, before the decalcification, the specimen was fixed with a Bouin's solution.
Preferably, the specific conditions of the fixation and decalcification steps are as follows: and (3) fixing the sample by adopting Bouin's solution for 16-24h, and decalcifying the sample for 30-60 min by using the bone marrow tissue decalcification solution.
The specimen to be inspected is placed into the Bouin's liquid for fixation after the specimen to be inspected is taken, the specimen to be inspected arrives at the laboratory, the fixation time reaches or even exceeds 16 hours basically, the specimen is completely fixed at the moment, glacial acetic acid and picric acid in the Bouin's liquid have the decalcification effect, although the effect is weak, the decalcification effect on bone marrow tissues is still certain after the specimen is soaked for 16 hours, then decalcification is carried out by using about 2% hydrochloric acid solution, only half an hour to one hour is needed, the decalcification process is relatively fast, and meanwhile, compared with other strong acids, the decalcification tissue is slightly damaged, the decalcification tissue can be softened by adding formaldehyde, so that slicing is facilitated, and the slicing effect is more ideal. The method has the advantages that the time of the manufacturing process is shortened, the manufactured section has the defects of clear tissue structure display, distinct nuclear texture contrast and large antigen destructiveness of relatively strong acid, the immunohistochemical staining cell membrane, cytoplasm and nucleus have excellent color development effect, the positioning is accurate, and the background is clean.
Further, after the fixation and decalcification steps are completed, dehydration, embedding, slicing, HE staining and/or immunohistochemical staining are sequentially performed.
The steps are conventional operations in the field, the fixing agent, the dehydrating agent, the clearing agent and the like used in the flaking process are reagents necessary for daily technical work, the slicing effect of the flaking process is good, the fixing, dehydrating, clearing, wax dipping and dyeing reagents used in the method can be repeatedly used for a plurality of times, and the reagent preparation and operation methods are simple and are worthy of clinical popularization and application.
Preferably, the dehydration is gradient dehydration, with the specific conditions: soaking in 10% formalin for 1-1.5 h, sequentially soaking in 75% alcohol for 1-1.5 h, soaking in 95% alcohol for 1-1.5 h, repeating for 1 time, soaking in anhydrous alcohol for 1-1.5 h, repeating for 2 times, soaking in xylene for 30-50 min, repeating for 2 times, soaking in paraffin for 1-1.5 h, and repeating for 2 times.
More preferably, the immersion is performed twice in 10% formalin → 1h in 75% alcohol → 1h in 95% alcohol → 2(× 2 means that the operation is performed twice, that is, after the immersion in 95% alcohol is performed for 1h, the immersion liquid is replaced, and the immersion is performed in 95% alcohol for 1h) → immersion in anhydrous alcohol 1h × 3(× 3 means that the operation is performed three times) → immersion in xylene 40min × 3 → immersion in paraffin 1h × 3.
HE staining procedure: baking 3-micron slices at 80 ℃ for half an hour → dewaxing agent 10min → 3 → absolute ethyl alcohol 3min → 95% ethyl alcohol 3min 2 → 75% ethyl alcohol 3min → flowing water washing 2min → hematoxylin 6min, flowing water washing 2min → 4% hydrochloric acid alcohol differentiation for 3-5 s, flowing water washing 2min → 1% ammonia water rewet for 10s, flowing water washing 2min → eosin 3-5 s, flowing water washing → gradient alcohol dehydration, and blow-drying the sealing piece to be detected.
Immunohistochemical staining procedure: (1)2 μm slices are baked at 80 deg.C for half an hour, and dewaxed to water conventionally; (2) putting the slide into a preheated repairing instrument, and operating a repairing program; (3) washing with distilled water for three times; (4) marking by an immunohistochemical oil pen; (5) and (6) loading on a machine. 3% of hydrogen peroxide, primary antibody, reinforcing agent, secondary antibody and DAB color development programs are completed; (6) taking down the slide, and washing with running water; (7) hematoxylin (BASO hematoxylin) for 2min, and washing with running water; (8) differentiating by 0.25% hydrochloric acid alcohol for 3-5 s, and washing by running water; (9) warm water is used for carrying out bluing and washing with running water; (10) and (5) dehydrating by gradient alcohol, and drying the sealing piece for inspection.
Wherein, the repairing instrument is preferably preheated to 75 ℃ in advance, and the repairing condition is 96 ℃ for 25 min.
The invention provides a bone marrow tissue decalcifying liquid and a method for manufacturing a bone marrow tissue paraffin section, the bone marrow tissue specimen is decalcified by adopting the decalcifying liquid, the specimen pretreatment process is shortened, the report time is short, the more timely diagnosis is facilitated, and the obtained HE section shows that the cell morphology is well preserved and the tissue structure is clear; the immunohistochemical staining section shows that the staining effect is excellent, the positioning is accurate, and the background is clean; and the sample does not have the phenomenon of excessive decalcification.
Drawings
FIG. 1 is a paraffin section (20X 10) of HE-stained bone marrow tissue in example 1 of the present invention;
FIG. 2 is a paraffin section (40X 10) of HE-stained bone marrow tissue in example 1 of the present invention;
FIG. 3 is a paraffin section (Bcl-6, 40X 10) of immunohistochemically stained bone marrow tissue according to example 1 of the present invention;
FIG. 4 is a paraffin section (CyclinD1, 40X 10) of immunohistochemically stained bone marrow tissue according to example 1 of the present invention;
FIG. 5 is a paraffin section (20X 10) of HE-stained bone marrow tissue in comparative example 1 of the present invention;
FIG. 6 is a paraffin section (40X 10) of HE-stained bone marrow tissue in comparative example 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The reagents used in the following examples are commercially available unless otherwise specified.
The immunohistochemical instrument used in the following examples is an AutostainerLink48 semi-automatic immunohistochemical instrument, and the repairing instrument is a PT Link immunohistochemical pretreatment system, all of which are products of DAKO corporation.
Example 1
This example provides a bone marrow tissue decalcifying liquid, which comprises hydrochloric acid (mass fraction is 2.34%), formaldehyde and distilled water at a volume ratio of 2:10:88, and has a pH of 1.35.
The embodiment also provides a method for manufacturing the paraffin section of the bone marrow tissue, which comprises the following specific steps:
1. fixation-decalcification: the specimen (cylindrical bone marrow tissue about 0.5-1 cm long) is fixed with Bouin's solution for 16-24h, and then treated with the above-mentioned decalcifying solution for 30 min.
2. Gradient dehydration: 10% formalin 1h → 75% alcohol 1h → 95% alcohol 1h 2 → absolute alcohol 1h 3 → xylene 40min 3 → paraffin 1h 3.
3. And (6) embedding in paraffin.
4. HE staining: baking 3-micron slices at 80 ℃ for half an hour → dewaxing agent 10min → 3 → absolute ethyl alcohol 3min → 95% ethyl alcohol 3min 2 → 75% ethyl alcohol 3min → flowing water washing 2min → hematoxylin 6min, flowing water washing 2min → 4% hydrochloric acid alcohol differentiation for 3-5 s, flowing water washing 2min → 1% ammonia water rewet for 10s, flowing water washing 2min → eosin 3-5 s, flowing water washing → gradient alcohol dehydration, and blow-drying the sealing piece to be detected.
5. Immunohistochemical staining: (1)2 μm slices are baked at 80 deg.C for half an hour, and dewaxed to water conventionally; (2) putting the slide into a preheated repairing instrument, and operating a repairing program; preheating the repairing instrument to 75 ℃ in advance, wherein the repairing condition is 96 ℃ for 25 min; (3) washing with distilled water for three times; (4) marking by an immunohistochemical oil pen; (5) loading on a machine to finish the color development procedures of 3% hydrogen peroxide, primary antibody, reinforcing agent, secondary antibody and DAB; (6) taking down the slide, and washing with running water; (7) hematoxylin (BASO hematoxylin) for 2min, and washing with running water; (8) differentiating by 0.25% hydrochloric acid alcohol for 3-5 s, and washing by running water; (9) warm water is used for carrying out bluing and washing with running water; (10) and (5) dehydrating by gradient alcohol, drying the sealing piece and checking for detection.
FIGS. 1 and 2 show paraffin sections of HE-stained bone marrow tissue in this example, in which the morphology of the cells is well preserved and the tissue structure is clear. Fig. 3 and 4 are paraffin sections of immunohistochemically stained bone marrow tissues in this example, and it can be seen from the figures that the staining effect is excellent, the location is accurate, and the background is clean. The specimens of this example did not show excessive decalcification and no residual acid treatment step was required.
Comparative example 1
The comparative example provides a bone marrow tissue decalcification solution, which consists of hydrochloric acid (mass fraction is 2.34%), formaldehyde and distilled water in a volume ratio of 10:10: 80.
This comparative example also provides a method for preparing paraffin sections of bone marrow tissue using the above-described decalcifying solution, the procedure being the same as in example 1.
Results the HE stained paraffin sections of bone marrow tissue were shown in fig. 5 and 6 and were less effective than those of example 1, with indistinct nuclear mass and indistinct cytoplasmic nuclear contrast.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (8)
1. The bone marrow tissue decalcification solution is characterized by comprising hydrochloric acid, formaldehyde and distilled water, wherein the hydrochloric acid accounts for 1-3% of the bone marrow tissue decalcification solution by volume percentage, and the formaldehyde accounts for 9-11% of the bone marrow tissue decalcification solution by volume percentage.
2. The bone marrow tissue decalcifying liquid according to claim 1, wherein the hydrochloric acid is present at a concentration of 2% by volume of the bone marrow tissue decalcifying liquid, and the formaldehyde is present at a concentration of 10% by volume of the bone marrow tissue decalcifying liquid.
3. The bone marrow tissue decalcifying liquid according to claim 1 or 2, wherein the hydrochloric acid is diluted hydrochloric acid with a mass fraction of 2-3%.
4. A method for preparing a paraffin section of a bone marrow tissue, comprising the step of decalcifying a specimen with the bone marrow tissue decalcifying liquid according to any one of claims 1 to 3.
5. The method of claim 4, wherein the specimen is fixed with Bouin's solution before the decalcification.
6. The method for preparing paraffin sections of bone marrow tissues according to claim 5, wherein the fixing and decalcification steps are carried out under the specific conditions: and (3) fixing the sample by adopting Bouin's solution for 16-24h, and decalcifying the sample for 30-60 min by using the bone marrow tissue decalcification solution.
7. The method of claim 6, wherein the fixing and decalcification steps are followed by dehydration, embedding, sectioning, HE staining and/or immunohistochemical staining.
8. The method for preparing paraffin sections of bone marrow tissue according to claim 7, wherein the dehydration is a gradient dehydration, with the specific conditions: soaking in 10% formalin for 1-1.5 h, sequentially soaking in 75% alcohol for 1-1.5 h, soaking in 95% alcohol for 1-1.5 h, repeating for 1 time, soaking in anhydrous alcohol for 1-1.5 h, repeating for 2 times, soaking in xylene for 30-50 min, repeating for 2 times, soaking in paraffin for 1-1.5 h, and repeating for 2 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011614151.8A CN112816295A (en) | 2020-12-30 | 2020-12-30 | Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011614151.8A CN112816295A (en) | 2020-12-30 | 2020-12-30 | Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112816295A true CN112816295A (en) | 2021-05-18 |
Family
ID=75855505
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011614151.8A Pending CN112816295A (en) | 2020-12-30 | 2020-12-30 | Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112816295A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109425523A (en) * | 2017-09-05 | 2019-03-05 | 上海新培晶医学检验所有限公司 | Marrow specimen processing method, decalcifying Fluid and purposes |
| CN111238904A (en) * | 2020-03-17 | 2020-06-05 | 四川金域医学检验中心有限公司 | Bone marrow decalcification solution beneficial to immunohistochemical color development and use method thereof |
-
2020
- 2020-12-30 CN CN202011614151.8A patent/CN112816295A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109425523A (en) * | 2017-09-05 | 2019-03-05 | 上海新培晶医学检验所有限公司 | Marrow specimen processing method, decalcifying Fluid and purposes |
| CN111238904A (en) * | 2020-03-17 | 2020-06-05 | 四川金域医学检验中心有限公司 | Bone marrow decalcification solution beneficial to immunohistochemical color development and use method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 周艳华: "60Coγ射线照射促进小鼠骨髓脂肪化研究", 《暨南大学学报（医学版）》 * |
| 姚洪田 等: "骨髓活检标本固定液的选择", 《肿瘤学杂志》 * |
| 李锐等: "两种盐酸脱钙液对骨髓HE及免疫组化染色的影响", 《中国医刊》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103940658A (en) | Method for manufacturing paraffin-embedded tissue cell specimen | |
| CN103930761B (en) | Unmixing binary system for pre-processing the biological specimen of embedding | |
| CN106644656A (en) | Hematoxylin-eosin one-step dyeing method | |
| EP3035026A1 (en) | Composition for preparing biomaterial with excellent light-transmitting property, and use thereof | |
| CN105588943A (en) | Detection method for peripheral blood CTC (Circulating Tumor Cell) Her-2 gene of stomach cancer patient | |
| Wolfe | Tissue processing | |
| WO2014114009A1 (en) | Immunohistochemical quality control reference object and quality control method | |
| RU2536502C2 (en) | Cytological and histological fixing composition and staining method | |
| Golberg et al. | Application of automated immunohistochemistry in anatomical research: A brief review of the method | |
| CN109946278A (en) | Fluorescent dye DAPI carries out cell DNA to dye quantitative screening for cancer and diagnostic method | |
| CN103994913A (en) | Fixed-decalcifying fluid for bone marrow biopsy and paraffin section method of bone marrow biopsy tissue | |
| CN112816295A (en) | Bone marrow tissue decalcification liquid and method for preparing bone marrow tissue paraffin section | |
| CN106198996B (en) | Early diagnosis biomarker, kit and its application of minimal change nephrosis | |
| CN105548569A (en) | Detection method for peripheral blood VEGF of renal cancer patient | |
| CN113720669B (en) | Immunohistochemical quality control product constructed based on animal organs or tissues | |
| CN105624304A (en) | Method of flounder juvenile gonad mRNA in-situ hybridization | |
| CN109187147A (en) | A kind of dyeing of arterial wall ingredient and identification method | |
| CN108693010A (en) | A kind of plant tissue softening agent and application method | |
| Al-Refu | General methods in preparation of skin biopsies for haematoxylin and eosin stain and immunohistochemistry | |
| Jafari et al. | Diagnostic value of immunoperoxidase staining and immunofluorescence in the study of kidney biopsy specimens | |
| Gruber et al. | Basic staining and histochemical techniques and immunohistochemical localizations using bone sections | |
| CN105259001B (en) | A kind of paraffin section pre-treating method for tissue biopsy | |
| RU2408887C1 (en) | Method to produce histological preparations and compositions for its realisation (versions) | |
| Kiernan | Nuclear stains | |
| Nakahara et al. | Detection of activated mouse neurons with temporal resolution via dual c-Fos staining |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210518 |
|
| RJ01 | Rejection of invention patent application after publication |