Immunohisto

Friday 7th. Dec 2007

Today is the first day I stain my slides. What I did today

1. 5 slides all together - diltion: 1/50, 1/250, 1/500, one slide with Hx alone, one slide with secondary alone
2. Water bath - It took 15 minutes to increase 10 0 C.
3. Dewaxing - Hot plate 60 0 C - 15 minutes
4. Rehydration - Xylene 2x, Alcohol 100 % 2x, 90 % , 80%, 70%. 50 % not used.
5. Antigen retrieval solution in jar - heated earlier
6. Slides put into jar - temperature drop a bit when the cover was lifted up. Tempertaure pick up above 95 0 C in about 10 minutes. Slides in ARS for 30 minutes
7. Cooling down - 30 minutes
8. TBS - Shaker 3 min x 2
9. H2O2 - 10 minutes
10. TBS shaker 3 min x2
11. Primary AB - 1 hour
12. TBS Shaker 3 min x2
13. Secondary antibody - 30 minutes
14. TBS shaker 3 min x2
15. Chromogen DAB 10 minutes
16. TBS Shaker 3 min x2
17. wash with DH2O
18. Hematoxylin 30 s..drip only at sink
19. wash with running water
20. Dehydrate - 70-80-90-100-100-xylene-xylene
21. wipe dry
22. DPX
23. dry

Monday baru boleh angkat