|From:||yichao wu |
I was told that sucrose could dehydrate the tissue and protect from ice cystals forming. Yes I will try to avoid it next time.
Up to now,I get the impression that snap freezing is the most important, right? So, may I do as follows(Could I use -70C fridge instead of solid ice)?
1. Put fresh kidney biopsy specimen in a Hank's BBS at 4C and transfer to the lab.
2. Precool isopentane in -70C fridge in small vials for several hours. Immediately put specimen into isopentane for 30 seconds.
3. Precool chuck in -70C fridge.Put a drop of OCT on the top of chuck and immediately transfer the frozened specimen to OCT and let them all frozen for 1 minutes.
4. Transfer them to -18C cyrostat and wait for 5 minutes.
5. Start to sectioning.
Could you kindly make some comments on this procedure?