CN113588383A - HE staining method for liver pathological large section - Google Patents
HE staining method for liver pathological large section Download PDFInfo
- Publication number
- CN113588383A CN113588383A CN202110661328.8A CN202110661328A CN113588383A CN 113588383 A CN113588383 A CN 113588383A CN 202110661328 A CN202110661328 A CN 202110661328A CN 113588383 A CN113588383 A CN 113588383A
- Authority
- CN
- China
- Prior art keywords
- ethanol
- liver
- 5min
- staining
- xylene
- 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
- 238000000034 method Methods 0.000 title claims abstract description 52
- 230000001575 pathological effect Effects 0.000 title claims abstract description 45
- 210000004185 liver Anatomy 0.000 title claims abstract description 43
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 title claims abstract description 39
- 208000006990 cholangiocarcinoma Diseases 0.000 claims abstract description 5
- 238000003745 diagnosis Methods 0.000 claims abstract description 4
- 238000012752 Hepatectomy Methods 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 108
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 37
- 239000008096 xylene Substances 0.000 claims description 23
- 238000004043 dyeing Methods 0.000 claims description 18
- 238000010186 staining Methods 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 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 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 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 claims description 9
- 239000012188 paraffin wax Substances 0.000 claims description 9
- 230000018044 dehydration Effects 0.000 claims description 7
- 238000006297 dehydration reaction Methods 0.000 claims description 7
- 230000007170 pathology Effects 0.000 claims description 7
- -1 5min Chemical compound 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- 208000019423 liver disease Diseases 0.000 claims description 4
- 230000007935 neutral effect Effects 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- DZGCGKFAPXFTNM-UHFFFAOYSA-N ethanol;hydron;chloride Chemical compound Cl.CCO DZGCGKFAPXFTNM-UHFFFAOYSA-N 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 2
- 230000002980 postoperative effect Effects 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 238000007789 sealing Methods 0.000 claims 1
- 210000001519 tissue Anatomy 0.000 abstract description 36
- 210000005228 liver tissue Anatomy 0.000 abstract description 11
- 206010019695 Hepatic neoplasm Diseases 0.000 abstract 2
- 208000014018 liver neoplasm Diseases 0.000 abstract 2
- 238000003759 clinical diagnosis Methods 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 31
- 210000004027 cell Anatomy 0.000 description 12
- 206010028980 Neoplasm Diseases 0.000 description 11
- 201000011510 cancer Diseases 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 230000036571 hydration Effects 0.000 description 6
- 238000006703 hydration reaction Methods 0.000 description 6
- 208000005156 Dehydration Diseases 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 239000008399 tap water Substances 0.000 description 4
- 235000020679 tap water Nutrition 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 3
- 210000002808 connective tissue Anatomy 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 210000004738 parenchymal cell Anatomy 0.000 description 2
- 238000010827 pathological analysis Methods 0.000 description 2
- 238000007447 staining method Methods 0.000 description 2
- 208000035346 Margins of Excision Diseases 0.000 description 1
- 208000008385 Urogenital Neoplasms Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 210000002187 accessory nerve Anatomy 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 210000003484 anatomy Anatomy 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004791 biological behavior Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- UKZQEOHHLOYJLY-UHFFFAOYSA-M ethyl eosin Chemical compound [K+].CCOC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 UKZQEOHHLOYJLY-UHFFFAOYSA-M 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000001926 lymphatic effect Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 208000037964 urogenital cancer Diseases 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000003462 vein Anatomy 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
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
-
- 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
- G01N2001/302—Stain compositions
Abstract
The invention relates to the technical field of pathological sections, in particular to an HE staining method for a large pathological section of liver. Specifically, a large pathological section of the liver tissue with the size of more than 5cm for diagnosis and scientific research is prepared by adopting the HE staining method provided by the invention based on the large-scale hepatectomy tissue of the hepatoportal cholangiocarcinoma. The large pathological liver section prepared by the HE staining method provided by the invention can display the microenvironment around the liver tumor and the relationship between the liver tumor and the surrounding tissues in a panoramic way, and is used for comprehensive clinical diagnosis and scientific research.
Description
Technical Field
The invention relates to the technical field of pathological sections, in particular to an HE staining method for a large pathological section of liver.
Background
The use of histology on large sections of pathology began in the 20 th century in the 70's brain histopathology. The reason for using pathologically large sections is to visualize the normal tissue of the brain and its structure and changes, which can correlate macroscopic and microscopic features and abnormalities of the brain with clinical manifestations, including providing information for imaging techniques emerging at the time.
The application of histopathology on large sections of urogenital cancer specimens is for the same reason as that of neuropathologists. For example, the use of large-slice histology is important to understand the normal anatomy of the prostate, especially for pathologists and radiologists who are trained to improve the "clinical view" of the clinician before diagnosis by correlation with radiographic images.
The precise information that a pathologist can define and forward to a clinician by the pathology gross section technique is: diagnostic and prognostic information, including tumor multifocal and location, histological type and variation, grade, TNM staging, including surgical margin status (R) and Lymphatic Vascular Infiltration (LVI), and tumor volume and size. Other features of clinical significance and other features of potential clinical significance can also be known by the technique of pathological gross section.
The liver pathological large section technology has great effect on the research of liver malignant tumor. The technology can be used for observing the microenvironment around the tumor in a panoramic way, positioning the relation between the tumor and surrounding tissues and the biological boundary of the tumor, but at present, no research-type paper report of systematic liver pathology large sections exists internationally.
HE staining of liver tissue is the key of the whole preparation process of pathological large liver sections. The liver is the largest parenchymal organ of the abdominal cavity, in addition, the number of parenchymal cells of the liver is large, the Glisson system in the liver is complicated and complicated, the shapes of all systems and cells in the liver need to be observed simultaneously, the steps and the difficulty of HE staining are greatly different from those of other tissues, and the high requirements on the quality of HE staining are provided.
Disclosure of Invention
In order to solve the problem that the conventional HE staining is not suitable for preparing pathological large sections, the invention provides an HE staining method suitable for liver pathological large sections. The invention aims to provide an HE staining method for preparing pathological liver tissues of pathological sections of more than 5cm, which is favorable for application in postoperative pathological diagnosis and scientific research of liver diseases.
In a first aspect, the present invention provides an HE staining method for a pathological large liver section, wherein a dewaxing method in the prior art cannot be applied to dewaxing a large specimen tissue because the wax content of the large specimen tissue of more than 5cm of a liver is high, and therefore, in the HE staining method, the hydration dewaxing treatment comprises:
xylene (I), 10min,
xylene (II), 10min,
xylene (III), 5 min;
the xylene removal treatment used:
100% ethanol (I), for 5min,
100% ethanol (II), 5min,
95% ethanol for 5 min;
80% ethanol, 5 min.
Because the liver tissue specimen is larger, if hydration treatment in the conventional dyeing step is adopted, more cells in the large liver tissue have poor coloring effect due to over expansion, and observation is influenced; and the volume of the large tissues of the liver expands rapidly, which can cause the surface of the tissues to generate concave-convex and never cause the failure of digital scanning imaging. In the hydration of large tissues, the use of three-stage alcohol concentration gradients and increased alcohol exposure times are essential.
Dyeing and differentiating treatment by using 1% hydrochloric acid for 20-40 s;
eosin staining is 0.5% aqueous eosin staining for 1-3 min.
In the dyeing process, the reason for using the aqueous eosin solution is that after dewaxing and xylene removal treatment, large tissues of the liver are hydrated, the combination degree of the alcohol-soluble eosin in water is poor, the alcohol must be quickly added after dyeing is finished, and if the dyeing is slow, the color is dropped.
In the HE staining method provided by the invention, the staining time of hematoxylin is 8-12 min.
In the HE dyeing method provided by the invention, the time for returning blue by using 1% ammonia water is 30 s.
The HE dyeing method further comprises dehydration treatment after dyeing, wherein the dehydration and transparent treatment after dyeing comprises the following steps:
80% ethanol, 3min, 95% ethanol, 5min, anhydrous ethanol (I), 5min, anhydrous ethanol (II)5min, xylene (I), 5min, and xylene (II), 5 min.
In the HE staining method provided by the invention, the pathological large tissue of the liver is a large-scale hepatectomy tissue of the hepatoportal cholangiocarcinoma.
Specifically, as a specific embodiment of the present invention, the HE dyeing method is:
(1) xylene (I), 10min, xylene (II), 10min, xylene (III), 5 min;
(2) 100% ethanol (I), 5min, 100% ethanol (II), 5min, 95% ethanol, 5min, 80% ethanol, 5min, washing with running water, 2min, and distilled water for 2 min;
(3) staining with hematoxylin for 10min, and washing with running water for 3 min;
(4) washing with 1% hydrochloric acid ethanol for 30s for 1 min;
(5) returning blue with 1% ammonia water for 30s, and washing with running water for 1 min;
(6) dyeing with 0.5% water-based eosin solution for 1-3min, and washing with running water for 1 min;
(7) 80% ethanol, 3min, 95% ethanol, 5min, anhydrous ethanol (I), 5min, and anhydrous ethanol (II) for 5 min;
(8) xylene (I), 5min, xylene (II), 5 min.
In a second aspect, the invention provides a method for preparing a large liver pathological tissue section, wherein after the liver pathological tissue is stained by the HE staining method, the large liver pathological tissue section is sealed by neutral gum, so that bubbles are reduced as much as possible, and the large stained liver pathological tissue section is obtained.
In a third aspect, the present invention provides a paraffin section for liver pathology, wherein the paraffin section is used for staining the pathological tissues of the liver by using the above HE staining method; the paraffin section is a liver pathological section of more than 5 cm.
According to the understanding of the skilled person, the invention also claims the application of the above HE staining method or the above preparation method or the above paraffin section in the pathological diagnosis after liver disease operation and the study of hepatoportal cholangiocarcinoma.
The invention has the beneficial effects that:
(1) the invention can fully dewax the tissue of a large sample with the size of more than 5cm by increasing the acting time of the dimethylbenzene, so that the tissue fully enters a water-soluble state;
(2) in the hydration process, the action time of 100 percent alcohol is increased, and the long-time hydration steps of 95 percent and 80 percent alcohol are also increased, so that the concentration and the action time of the alcohol used for treatment are increased, the hydration gradient of liver tissues is more moderate, and the cells are fully prevented from over-expansion;
(3) the aqueous eosin solution adopted by the invention is easy to combine with hydrated large specimen tissues, and the staining is more stable in HE staining of large specimens;
(4) the invention provides a method for staining pathological tissues of liver, wherein a large pathological section prepared by the staining method is suitable for the diagnosis-level research of liver diseases, the cell nucleuses of various cells in the pathological section are clearly displayed, and the cells and tissues are uniformly stained;
(5) the large pathological liver section prepared by the HE staining method can clearly distinguish the accessory nerves, arteries, veins and fibrous connective tissues of the liver under a microscope, and is favorable for observing the biological behavior of tumors.
Drawings
FIG. 1 is a diagram of pathological tissues of liver used in example 1 of the present invention.
FIG. 2 is a diagram of atypical cells obtained by microscopic observation of a large pathological section in example 1 of the present invention.
FIG. 3 is a diagram of parenchymal hepatocytes obtained by microscopic observation of a large pathological section in example 1 of the present invention.
FIG. 4 is a microscopic view of a large pathological section in comparative example 1 of the present invention.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. It is intended that all modifications or alterations to the methods, procedures or conditions of the present invention be made without departing from the spirit and substance of the invention.
Unless otherwise specified, test materials, reagents, instruments and the like used in the examples of the present invention are commercially available; all technical measures in the examples of the present invention are conventional measures well known to those skilled in the art, unless otherwise specified.
Example 1
This example provides a method for HE staining of pathological tissues to prepare large pathological sections of 5cm or more.
First, HE dyeing pretreatment
As shown in figure 1, the liver tissue after operation is soaked in 10% formalin 10 times of the specimen for 24 hours, and then the material is obtained by using a material-obtaining knife in a continuous radial direction with the cross section of 8-10mm thickness (the plane parallel to axial abdominal enhancement CT). And then, secondarily taking the specimen, and under the guidance of an embedding frame, secondarily taking the specimen by slicing at the same angle around the human body to ensure the flatness of the large specimen, wherein the tissue thickness of each slice is controlled to be 4-5 mm. Fully soaking the tissues in 10% neutral formalin which is 10 times of the tissues, fixing for 24 hours again, and entering an HE staining stage after the dehydration machine baking procedure is completed.
Secondly, the HE dyeing method comprises the following steps:
after coloring is finished, the invention adopts 80 percent, 95 percent and absolute ethyl alcohol gradient alcohol to dehydrate in 18 to 21 steps, and controls dehydration time, so that dehydration process is more moderate, and pathological observation is effectively prevented from being influenced by rapid shrinkage of cells of large slices. Meanwhile, the occurrence of unevenness on the surface of the tissue is prevented, so that the digital imaging fails.
And finally, using an Olympus VS200 panoramic digital scanner to read the film after scanning is finished.
After the section is read by a pathologist, the whole liver tissue of the large liver tissue section prepared by the embodiment is uniformly colored, no obvious color difference is seen, hepatocellular carcinoma cells are clearly displayed after further amplification, abnormal cells (shown in figure 2) can be clearly distinguished after 200 times of amplification, blood vessels, nerves and fibrous connective tissues around the cancer tissue can be clearly observed, and the microenvironment around the cancer and the relation with the surrounding tissues can be accurately observed. The parenchymal hepatocytes and nuclei were clearly shown (see FIG. 3). The HE staining method can be universally applied to large pathological tissues of the liver.
After the section is read by a pathologist, the whole liver tissue of the large liver tissue section prepared by the embodiment is uniformly colored, no obvious color difference is seen, hepatocellular carcinoma cells are clearly displayed after further amplification, abnormal cells can be clearly distinguished after 200 times of amplification, blood vessels, nerves and fibrous connective tissues around the cancer tissue can be clearly observed, and the microenvironment around the cancer and the relation between the microenvironment around the cancer and the surrounding tissues can be accurately observed. Parenchymal hepatocytes and nuclei were clearly shown. The HE staining method can be universally applied to large pathological tissues of the liver.
Comparative example 1
In order to obtain a liver pathological section with clear staining of more than 5cm, the invention carries out a plurality of attempts in different improvement directions on the staining method, and the comparative example provides an HE staining method for a large liver pathological tissue, and is different from the example 1 in that the HE staining method adopted by the comparative example is as follows:
(1) 3min of dimethylbenzene;
(2) 3min of dimethylbenzene;
(3) 3min of dimethylbenzene;
(4) absolute ethyl alcohol for 3 min;
(5) 95% ethanol for 3 min;
(6) 95% ethanol for 3 min;
(7) 80% ethanol for 3 min;
(8) 70% ethanol for 3 min;
(9) 1min tap water;
(10) 7min of hematoxylin;
(11) tap water lmin;
(12) 0.5-1% hydrochloric alcohol < 75% alcohol > differentiation solution for 30 s;
(13) 1min tap water;
(14) returning the 0.5-1% ammonia water to the blue solution for 30 s;
(13) tap water lmin;
(14) 0.5% alcohol < 80% ethanol > soluble eosin 30 s;
(15) 70% ethanol for 30 s;
(16) 80% ethanol lmin;
(17) 95% ethanol for 2 min;
(18) 95% ethanol for 2 min;
(19) absolute ethyl alcohol for 1 min;
(20) 3min of dimethylbenzene;
(21) 3min of dimethylbenzene;
(22) 3min of dimethylbenzene;
(23) and (5) entering a mounting machine for mounting neutral gum.
The microscopic observation picture of the large section obtained by the comparative example is shown in figure 4, and the staining program of the comparative example can obtain the large pathological section of the liver which is not uniformly stained, can not accurately distinguish the cell nucleus of various cells, and declares the staining failure.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. An HE staining method for pathological large liver sections is characterized in that the HE staining method comprises the following steps:
xylene (I), 10min,
xylene (II), 10min,
xylene (III), 5 min;
the xylene removal treatment comprises the following steps:
100% ethanol (I), for 5min,
100% ethanol (II), 5min,
95% ethanol for 5 min;
80% ethanol for 5 min;
dyeing and differentiating by treating with 1% hydrochloric acid ethanol for 20-40 s;
eosin staining is 0.5% aqueous eosin staining for 1-3 min.
2. The HE staining method according to claim 1, wherein the time for hematoxylin staining in the HE staining method is 8 to 12 min.
3. The HE staining method of claim 2, wherein 1% ammonia is used to turn blue for 30 s.
4. The HE dyeing method according to claim 3, further comprising a post-dyeing dehydration and clearing process, the post-dyeing dehydration and clearing process being:
80% ethanol, 3min, 95% ethanol, 5min, anhydrous ethanol (I), 5min, anhydrous ethanol (II)5min, xylene (I), 5min and xylene (II), 5 min.
5. The HE staining method of any of claims 1-4, wherein the pathological large tissue of the liver is a large hepatectomy tissue of the hepatoportal cholangiocarcinoma.
6. The HE staining method of claim 5, wherein the HE staining method is:
(1) xylene (I), 10min, xylene (II), 10min, xylene (III), 5 min;
(2) 100% ethanol (I), 5min, 100% ethanol (II), 5min, 95% ethanol, 5min, 80% ethanol, 5min, washing with running water, 2min, and distilled water for 2 min;
(3) staining with hematoxylin for 10min, and washing with running water for 3 min;
(4) washing with 1% hydrochloric acid ethanol for 30s for 1 min;
(5) returning blue with 1% ammonia water for 30s, and washing with running water for 1 min;
(6) dyeing with 0.5% water-based eosin solution for 1-3min, and washing with running water for 1 min;
(7) 80% ethanol, 3min, 95% ethanol, 5min, anhydrous ethanol (I), 5min, and anhydrous ethanol (II) for 5 min;
(8) xylene (I), 5min, xylene (II), 5 min.
7. A method for preparing a large section of a liver pathological tissue, which comprises staining the liver pathological tissue by the HE staining method according to any one of claims 1 to 6, followed by sealing with a neutral gum to minimize air bubbles and obtaining a large section of the stained liver pathological tissue.
8. A paraffin section for liver pathology, wherein the paraffin section is stained for the liver pathology using the HE staining method of any one of claims 1 to 6; the paraffin section is a liver pathological section of more than 5 cm.
9. Use of the HE staining method according to any one of claims 1 to 6 or the preparation method according to claim 7 or the paraffin section according to claim 8 for the diagnosis of post-operative pathologies of liver diseases.
10. Use of the HE staining method according to any one of claims 1 to 6 or the preparation method according to claim 7 or the paraffin section according to claim 8 in the study of hepatoportal cholangiocarcinoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110661328.8A CN113588383A (en) | 2021-06-15 | 2021-06-15 | HE staining method for liver pathological large section |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110661328.8A CN113588383A (en) | 2021-06-15 | 2021-06-15 | HE staining method for liver pathological large section |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113588383A true CN113588383A (en) | 2021-11-02 |
Family
ID=78243850
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110661328.8A Pending CN113588383A (en) | 2021-06-15 | 2021-06-15 | HE staining method for liver pathological large section |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113588383A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150050689A1 (en) * | 2012-03-06 | 2015-02-19 | Roberto Gerigk | Formalin-Free Fixation Agent For Histological Stains of Tissue Samples |
| CN106644656A (en) * | 2017-01-06 | 2017-05-10 | 杨海玉 | Hematoxylin-eosin one-step dyeing method |
| CN112014192A (en) * | 2020-09-04 | 2020-12-01 | 贵州大学 | HE staining method for paraffin section of northern Guizhou goat ovary tissue |
| CN112798381A (en) * | 2021-02-03 | 2021-05-14 | 陕西省人民医院 | Multi-view tumor tissue pathological section and preparation method thereof |
-
2021
- 2021-06-15 CN CN202110661328.8A patent/CN113588383A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150050689A1 (en) * | 2012-03-06 | 2015-02-19 | Roberto Gerigk | Formalin-Free Fixation Agent For Histological Stains of Tissue Samples |
| CN106644656A (en) * | 2017-01-06 | 2017-05-10 | 杨海玉 | Hematoxylin-eosin one-step dyeing method |
| CN112014192A (en) * | 2020-09-04 | 2020-12-01 | 贵州大学 | HE staining method for paraffin section of northern Guizhou goat ovary tissue |
| CN112798381A (en) * | 2021-02-03 | 2021-05-14 | 陕西省人民医院 | Multi-view tumor tissue pathological section and preparation method thereof |
Non-Patent Citations (10)
| Title |
|---|
| 刘朝霞;徐丽;王军梅;杜江;李桂林;: "冰冻切片技术在神经病理诊断中的应用", 现代医药卫生, no. 06 * |
| 向思敏;王德玉;: "病理诊断中病理技术HE染色的临床价值研究", 临床医药文献电子杂志, no. 26 * |
| 宋彬妤;: "对制作高质量病理标本的方法研究", 当代医药论丛, no. 23 * |
| 徐加誉;廖子龙;张培荣;: "病理技术HE染色在病理诊断中的应用效果评价", 黑龙江中医药, no. 05 * |
| 李慧玲;张文莲;赵英芳;: "改善HE染色技术提高病理石蜡切片制作质量的体会", 包头医学院学报, no. 06, pages 103 * |
| 林斯燕;: "改良HE染色在病理诊断中的临床应用探讨", 基层医学论坛, no. 07 * |
| 田玉旺;朱红艳;朱培双;邢宜;: "环保型染色套液在常规病理组织切片染色中的应用", 诊断病理学杂志, no. 10, pages 02 - 04 * |
| 邢延青;: "病理技术HE染色在病理诊断中的应用效果观察", 世界最新医学信息文摘, no. 42 * |
| 郑燕璇;王晓鸿;曹婉维;李孜孜;李巍;: "肾穿组织免疫荧光及HE染色方法的改良", 中国医学创新, no. 30 * |
| 郑菲;陈培琼;王玉环;: "HE染色在临床病理诊断中的应用研究", 中外医疗, no. 30 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI753448B (en) | Method for analyzing tissue specimens | |
| CN107727463B (en) | Preparation of super-thick tissue slice and method for reproducing three-dimensional morphological structure of tissue at cell level | |
| CN109000956A (en) | A kind of closely-spaced secondary serial sectioning production method of full tumor of breast | |
| US20200388032A1 (en) | Three dimensional histopathology imaging method and system thereof | |
| Wang et al. | Three-dimensional reconstruction of light microscopy image sections: present and future | |
| CN104977194B (en) | A method of addition graphene accelerates sample process | |
| US10908246B2 (en) | Method for three-dimensional reconstruction of fascicular structure of human peripheral nerve | |
| CN113588383A (en) | HE staining method for liver pathological large section | |
| Zhu et al. | A versatile vessel casting method for fine mapping of vascular networks using a hydrogel-based lipophilic dye solution | |
| Chen et al. | Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains | |
| CN113588384A (en) | Dehydration method of liver pathological large tissue | |
| CN109859304B (en) | Three-dimensional printing technology for establishing three-dimensional structure digital model outside cornea limbal tissue | |
| Bochner et al. | Bimodal magnetic resonance and optical imaging of extracellular matrix remodelling by orthotopic ovarian tumours | |
| CN110003951B (en) | Paraffin for pathological semi-thin section and preparation method thereof | |
| KR101645085B1 (en) | A novel scanning electron microscopic tissue processing method for diagnosis of malignant mesothelioma using paraffin embedded pathologic specimen | |
| CN113138110A (en) | Three-dimensional reconstruction method for pathological picture | |
| CN110296935A (en) | A kind of entity tumor three-dimensional pathological diagnosis method | |
| CN105911281A (en) | Method for early warning breast cancer metastasis based on ZEB-1 and ZEB-2 as markers | |
| US20210231540A1 (en) | Method for preparation of tissue sections | |
| RU2799815C1 (en) | Method of macroscopic panoptic visualization of lesions and calculation of the volume of damaged myocardium when modeling a heart contusion | |
| WO2013088693A1 (en) | Staining kit and staining method | |
| Ding et al. | Rapid en-bloc hematoxylin-eosin staining for human lung cancer tissue for fluorescence micro-optical sectioning tomography | |
| CN111474120B (en) | Polarization imaging detection method for cells on non-deparaffinized-non-stained section | |
| CN116246019B (en) | 3D reconstruction method, device, equipment and medium for pathological section | |
| Lin et al. | Large tissue archiving solution for multiplexed labeling and super-resolution imaging |
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 |