[Histonet] Paraffin processing of muscle

[Jo Dee Fish]

Hi histonetters,

Sorry, I forgot to put anything in the subject with my first post, so I am
resubmitting it with a subject line!

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