Table 1 Protocol to deparaffinize the sections of FFPE tissue
1. Fresh xylenes (to deparaffinize the sections)-5 min | 1 |
2. Fresh xylenes-5 min | 2 |
3. 100% ethanol-30 sec | |
4. 95% ethanol-30 sec | |
5. 70% ethanol-30 sec | |
6. Deionized water-30 sec | |
7. Mayer's Hematoxylin (10%)- 30 sec | |
8. Deionized water-rinse 15 sec (x 2) | |
9. 70% ethanol-30 sec | |
10. Eosin Y (10%) - 15 sec | |
11. Deionized water-30 sec (x 2) | |
12. 3% glycerol in deionized water-30 sec | |
13. After removing the slide from the 3% glycerol step, shake the slide in the air to remove the layer of glycerol/water. | |
14. The next 5–10 mins are the optimal time for microdissection. The tissue is dry, but retains a soft consistency. If the dissection takes more than a few minutes, the tissue will become increasingly brittle and the dissected fragments may be repelled as the needle is brought in proximity to the tissue. If the tissue becomes overly dry, re-soak in the 3% glycerol/water solution for 1–2 minutes. |