[Histonet] Re: Cryosections from formalin fixed tissues

[kotresh mattur]

 Hi dear all,
 We r trying to get cryosections from 10% NBF fixed tissues for Oil Red 'O' staining. 

 First we   kept the fixed tissues for washing under tap water for overnight and then tried cryosectioning where we didnot get the quality sections

 Later we tried by keeping the tissues under 20% sucrose solution under refrigeration overnight(as mentioned by Gayle callis )and then we got better quality sections compared to the above said method. 

still we have few querries, can anybody give us ideal procedure for the same with your practical experiences? 

1. How does sucrose acts in preserving the tissues? i have read that it acts as cryoprotectant but i want to know how exactly it helps 

2.If we r keeping the 10% NBF fixed tissues in 20% sucrose solution then for how many days the tissues can be kept in the same solution under ref.? If so does it affect or come in the way of Oil Red 'O' staining and evaluation? 

3.After taking cryosetions if i want to preserve the same tissues whether they can be placed back to  10% NBF ?

Thanks in advance 

Dr.K.Y.Mathur



Dr.K.Y.Mathur
Junior Scientist
Dept.of Preclincal Safety Evaluation
Discovery Research
Dr.Reddy's Laboratory Ltd.
Bollaram Road, Miyapur,
HYDERABAD-INDIA-49.
 
Ph No: 
040-23045439-exn 424,421,422,423
Cell :09391293129

__________________________________________________
Do You Yahoo!?
Tired of spam?  Yahoo! Mail has the best spam protection around 
http://mail.yahoo.com