[Histonet] digesting soft tissue

[Philip Oshel]

Gayle,

Nice post -- I'd forgotten about Biz. Works a treat. We've used it 
for high-resolution SEM after digesting away the soft tissue.

Phil

>This is actually a published method in J of Anatomy to produce museum
>specimens for teaching sometime in early 80's???
>
>Buy BIZ laundry detergent (or raid the family laundry supply!) make a 20%
>solution, heat to 80C, less than boiling.  Suspend bones in this hot
>solution (or suspend and bring solution to working temperature, don't
>boil!)using cheesecloth to retain small boney parts that can fall off and
>handle a slippery, hot specimen easily. BIZ enzyme detergent loosens soft
>tissue to the point you can push it off with gloved fingers.  It will take
>cartilage off but that was acceptable.  Wear a mask when weighing - BIZ
>dust is very irritating to nasal passages and also skin. Digestion takes
>several hours if bone is large, so test frequently to see if soft tissue is
>releasing.
>
>I have done formalin fixed bone, rinsed over night in running tap water to
>remove the formalin (NBF kills the enzyme!), then BIZ.  NBF fixed bone
>stablilizes the soft tissue attachments to make them more resistant to
>enzyme. Wear gloves, safety glasses and rinse off detergent before handling
>bone excessively, slippery, and irritates skin.  Push soft tissue off, use
>old tooth brush and a dull knife to do gentle scraping if needed. When
>finished, rinse bone well, then air dry.  The result is a BLEACHED, clean
>bone that doesn't smell bad - particularly when the digestion process is
>happening.  Fixed is less smelly, unfixed means you are cooking bone while
>enzyme works - not so pleasant - use a hood.
>
>Have done whole bovine calf heads, bovine vertebral columns, bones from
>hind and forelimbs;  rat and mouse heads, rat femurs/tibias.  Beware - the
>enzyme will digest any soft attachments IF you let bones digest too long.
>Bones will fall apart, then you have to be a good anatomist and glue
>(thicker superglue works) tiny parts back in their relative,
>interdigitating, tight positions - a tedious but educational procedure on
>how our boney parts go together.
>
>
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>
>
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-- 
Philip Oshel
Supervisor, BBPIC microscopy facility
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison,  WI  53706 - 1284
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)