The paraffin section is the most widely used method in histological routine production techniques. Paraffin sections are not only used to observe the morphological structure of normal cell tissues, but also the main methods used by pathology and forensic science to study, observe and judge the morphological changes of cell tissues, and have been widely used in many other subject areas. in. In the teaching, the slice specimens observed under the light microscope are mostly prepared by paraffin sectioning. Living cells or tissues are mostly colorless and transparent, and there is a lack of contrast between various tissues and various structures within the cells. It is not easy to distinguish clearly under normal light microscope; tissue will soon die and produce tissue corruption after leaving the body. Loss of the original normal structure, therefore, the tissue should be fixed, paraffin embedded, sliced ​​and stained to avoid cell death, and can clearly identify its morphological structure.
Paraffin section HE staining (conventional HE staining) steps:
(1) xylene I: 5 to 10 min;
(2) xylene II: 5 to 10 min;
(3) Anhydrous ethanol I: 1 to 3 min;
(4) Anhydrous ethanol II: 1 to 3 min;
(5) 95% ethanol I: 1 ~ 3min;
(6) 95% ethanol II: 1 ~ 3min;
(7) 80% ethanol: 1 min;
(8) distilled water: 1 min;
(9) Sumu semen staining: 5 ~ 10min;
(10) Washing water to remove hematoxylin: 1 min;
(11) 1% hydrochloric acid-ethanol: 1 to 3 s;
(12) slightly washed: 1 ~ 2s;
(13) return to blue (with warm water or 1% ammonia, etc.): 5 ~ 10s;
(14) running water rinse: 1 ~ 2min;
(15) Distilled water washing: 1 to 2 min;
(16) 0.5% eosin staining: 1 ~ 3min;
(17) slightly washed distilled water: 1 ~ 2s;
(18) 80% ethanol: 1 to 2 s;
(19) 95% ethanol I: 2 to 3 min;
(20) 95% ethanol II: 2 to 3 min;
(21) Anhydrous ethanol I: 3 to 5 min;
(22) Anhydrous ethanol II: 3 to 5 min;
(23) carbolic acid-xylene: 3 to 5 min;
(24) xylene I: 3 to 5 min;
(25) xylene II: 3 to 5 min;
(26) xylene III: 3 to 5 min;
(27) Neutral gum seal.
Note: Items (12) and (13) above can be omitted, but the flushing time of (14) must be extended to 10-15 minutes (the nucleus shows clearer); (23) can be replaced by anhydrous ethanol, and the northern area can be omitted. .
Description: Paraffin sectioning methods include taking, fixing, washing and dehydrating, transparent, dipping wax, embedding, slicing and sticking, dewaxing, dyeing, dehydration, transparency, and sealing. It takes several days for a typical tissue to be made from a fixed specimen to a cover sheet, but the specimen can be stored for a long time as a permanent microscopic slide specimen.
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