[Histonet] muscle digest for sekeltal prep

From:"Boyce, Amanda (NIH/NIAMS)"

We are processing animals for skeletal preps that have been in 10% PFA for
quite some time (many months).  The first steps involved skinning and
eviscerating the embryos, followed by alcian blue staining.  We then put the
embryos in a trypsin digetion solution, which normally chews up the majority
of the muscle.  The rest of the muscle will be dissolved after the skeletons
are stained with alizarin red and put through a series of KOH washes.  The
problem is that because these animals are overfixed, the trypsin digest is
not sufficient.  Does anyone have any suggestions for a more vigorous
digestion?  Thanks for your help!  Amanda

Amanda Taylor Boyce, Ph.D.
Postdoctoral IRTA Fellow
Cartilage Biology and Orthopaedics Branch
National Institute of Arthritis and Musculoskeletal and Skin Diseases
National Institutes of Health
Bldg. 13, Rm. 3W17
Bethesda, MD 20892-5755
Phone: 301-451-6860
Fax: 301-480-4315
Email: boycea@mail.nih.gov

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