1. Direct observation of the morphological characteristics of living cells in an inverted microscope
2. Cell staining check:
a) placing the sterile slide in a cell culture dish, adding the cell suspension and culturing;
b) After the cells are full of slides, take them out, wash them with PBS, then place them on a glass slide, and let them dry naturally;
c) fixed with PBS/methanol (1:1) for 15 minutes;
d) discard the fixative and add suitable staining solution;
e) After dyeing, wash with water and dry in air.
f) After drying, it is transparent with xylene, and the optical resin is sealed and observed.
Second, immunohistochemical identification
1. Place the sterile coverslip in a cell culture dish and inoculate the cell suspension in a Petri dish for cell climbing;
2. Wash the specimen 3 times in PBS for 1 minute each;
3. Fix 4% paraformaldehyde for 15 minutes;
4. Place in the air to dry;
5. Wash the specimen 3 times in PBS for 2 minutes each;
6. Incubate 0.5% Triton X-100 once for 20 minutes;
7. Wash the specimen 3 times for 2 minutes each time;
8. Incubate for 15 minutes with 3% H 2 O 2 ;
9. DPBS cleans the specimen 3 times for 2 minutes each time;
10. Block serum incubation (5% normal secondary antiserum DPBS solution for 20 minutes)
11. Incubate with primary antibody at 4 ° C overnight or 37 ° C for 60 minutes;
12. Wash the specimen 3 times for 5 minutes each time;
13. Incubate the secondary antibody working solution (wet box) at 37 ° C for 30 minutes;
14. Wash the specimen 3 times for 5 minutes each time;
15. Liquid C (wet box) 37 ° C for 30 minutes;
16. Wash the specimen 3 times in PBS for 5 minutes each;
17. Develop color with a color developing solution;
18. Wash the distilled water twice for 1 minute;
19. Hematoxylin counterstaining for 1 minute;
20. Wash in clean water for 30 minutes;
21. Observe the optical resin after sealing.
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