[Histonet] (no subject)

[Jo Dee Fish]

Hi histonetters,
I have a question for you about muscle processing for paraffin embedding.
Starting from fixation to embedding, can you tell me the best and most
successful way to do this?  I have processed leg muscle (gastroc and soleus)
after initial overnight fixation in 3% paraformaldehyde using the following
schedule on our Excelsior:
 
Alcohols 1 through 6                        45 minutes each
Xylenes substitute 1 through 3           1 hour each
1st Paraffin @ 62 degrees                    1.5 hours
2nd Paraffin                                          1.5 hours
3rd Paraffin                                           2 hours
 
I know the paraffin sounds long, but we usually process fatty tissue and
brain that seems to require a longer infiltration time.
 
What do you think, am I missing the mark?  Our muscle seems to have a little
more space surrounding the bundles than the investigator had at their old
institute.
 
I really appreciate any help you can give.
 
Take care,
Jo Dee Fish
 
Jo Dee Fish

Research Technologist III

Gladstone Institute of Cardiovascular Disease 

Telephone: (415) 734-2567

Fax: (415) 355-0230

E-mail: jfish <@t> gladstone.ucsf.edu

Mailing address:  

The J. David Gladstone Institutes

1650 Owens Street

San Francisco, CA 94158