[Histonet] Re: Histonet Digest, Vol 106, Issue 35

[Galina Deyneko]

Dear Colleagues
I would like to ask again about cell block preparation. of cause I found number of answer on the histonet site and sorry that I ask again.
The cells what I prepare look distorted. Short protocol: 30 minutes of fixation in 10 %NBF, centrifuge  1000 rmr, wash in PBS, centrifuge, re-suspend in histogel and processed in Thermo Shandon with following program: Ethanols70, 70,80, 95,95,100,100,100,Xylenes 3 changes, Wax 3 changes - 1 hour in each station. I also tried short protocol - 30 minutes in each station.
I also would like to try to do OCT embedding to avoid processing anparaffinin embedding. Should I fix the cells before OCT embedding or not? Can I re-suspend directly in OCT or still need to embed in Histogel  first and freeze in OCT Histogel block. 
Please could you share your protocols and give me some hints.
Thank you and good weekend.


Galina Deyneko
Novartis, Cambridge, MA
 
617-871-7613 w