processing rat muscle

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From:"Yandl, Emily" <> (by way of histonet)
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Hi all!

I'm hoping someone can help me with this problem.  Our lab is working on a
project for a researcher who is looking at rat sciatic nerve along with the
associated muscle.  So far we have tried two different processing procedures
with less than satisfactory results.  Following both processing procedures,
upon cutting, the muscle appears very glassy and yellowish, like amber.
After the longer cycle, full sections were able to be cut after soaking, but
the sections exploded after being placed on the water bath.  After the
shorter cycle, the muscles held together better on the waterbath, however
developed ragged holes in the center of the tissue.  The pieces of muscle
approx. 2cm x 1.5 cm x 0.6 cm in dimension, and were processed on a Shandon
Citadel 2000 processor.

If anyone has any processing schedules that work for them or ideas of why
this isn't working, I would love to hear your comments.  The processing
schedules used are listed below.

The longer schedule is:
at least 48 hrs. 10% NBF fixation
2--30 min changes PBS
1--1hr 70% Alc
2--1hr changes 90% Alc
2--1hr changes 100% Alc
3--1.5 hrs changes in Xylene
2--2 hr changes in paraffin

The shorter schedule is:
at least 48 hrs in 10% NBF fixation
2--30 min changes in PBS
1--45 min change in 70% Alc
2--45 min changes in 90% Alc
2--45 min changes in 100% Alc
3--45 min changes in Xylene
2--2 hr changes in paraffin.

Thanks in advance for any help,

Emily Yandl
Pre-Clinical Biology
Genzyme Corp.
1 Mountain Rd.
Framingham, MA  01701
(508) 872-8400 x23623

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