I Got acceptable frozen sections with L N2 and wait for your comments

From:yichao wu

Dear Tony and Callis, Thank you very much.

Yesterday I got better results. I did as follows,

1) Transfer the renal biopsy specimen to lab within 0.9% NaCl moistened gauze

2) Embed the specimen on a chuck with OCT at room temperature. (The chuck was precooled to about -20C)

3)Put the chuck into liquid N2 directly for 1 minutes (Does it acceptable?)

4)Take out the chuck and put it into a holder(-33C) in the cyrostat and wait for 15 minutes

5)Start to section(-29C)

The morphology is acceptable.I should say, much better than before.

Dr. Henwood, I wish to try -18C as the sectioning temperature as you suggested for renal biopsy specimen. BTW,I wonder before the step 2, is it advisable to add PFA fixation and sucrose incubation? In order to improve the morphology further.

Dr. Callis,Thank you very much for the details about LCM.As you know, I have really little knowledge about cryostat.So I even could not differentiate cryomold and chuck (partly due to the language barrier ).I would like to contact a local supplier myself.Since ours was from Bright instrument OTF 5030, I contact Bright instrument in UK twice by email.But I have not got any replies.I am very anxious, really.

Thank you all.

Yichao WU

Jinling Hospital





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