[Histonet] digesting soft tissue

Thu Mar 18 15:27:37 CST 2004

Found these in the archives.  I thought I remembered this coming up
before. 

Enjoy.

Ross M Stapf
Histopathology Manager
Baylor University Medical Center
3500 Gaston Ave.
Dallas, TX 75246
214-820-2465
214-820-4110 fax
RossS <@t> baylorhealth.edu

To macerate tissue away, use Biz detergent (local grocery store)- an
enzyme
detergent.  Make a 20% solution and heat bone at 80C until muscle comes,
slides off bone surfaces. The enzyme is the kicker here, eats away at
protein attachments.  It bleachs bones as well, smells better than just
boiling bones.  After Biz treatment, rinse bones well with running tap
water - let them air dry. 

It is a good idea to fix bones with NBF or 70% alcohol, then rinse well
overnight to get rid of formalin - you don't want to inactivate enzyme
in
detergent. Also, fixed bone holds together better (cartilage is removed
by
BIZ!) and if bones (or parts of same) separate, glue with Duco Cement,
Superglue, or a glue gun after total drying.  Nice way to learn bone
anatomy by putting it back together. 

This method came from Journal of Anatomy, many moons ago, 1980's - to
prepare skulls for gross specimen study.

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis <@t> montana.edu

This is for those who would like something whose odor befits its work.

I think Gayle Callis' BIZ might be better, but Gee Whiz folks, we're
talking
about Gross Anatomy here!  These are procedures that one used to be
forced
to carry out on the roof, or out in an open field, away from civilized
folk.
So, I thought I would contribute a recipe from an old, VERY reputable,
source.

"Liquid Soap". Reighard and Jennings, "Anatomy of the Cat" [Oops! Sorry
kitty!], 1935, Holt Reinhart

Soft water		2000ml ("Soft" is not a scientific term!)
Strong ammonia	150ml  (for glazed eyes, opaque NICtitating membranes
and,
of course, alkalinity)
KNO3			12g (for suppression of something? John and
Abigail?)
Hard soap (brown)	75g  (We used to save the shavings of the brown
soap
used in the local Cub Scout carving tutorials.  We liked the brown soap,
because it WAS harder than Ivory!  This soap is still available in a
double
bar with attractive white wrapping with pink printing.  You can't tell a
soap by its cover!)

Heat the mixture to boiling until homogeneous.
Immerse the bone and boil for 40 min
Pour off the liquid and renew it.
Boil another 30min or until the soft parts come away easily.
[Boil too long and the epiphyses will separate.]
The bone can be rinsed in water.
The bone can be defatted, after drying, in bnenzene/zylene.  

Cheers (it almost feels like Friday),

Fred Monson

Frederick C. Monson, PhD
Center for Advanced Scientific Imaging
Mail to Geology
West Chester University of Pennsylvania
Schmucker II Science Center, Room SS024
South Church Street and Rosedale Avenue
West Chester, PA, 19383
Phone:  610-738-0437
eMail:  fmonson <@t> wcupa.edu

-----Original Message-----
From: Kim Merriam [mailto:kmerriam <@t> tktx.com]
Sent: Tuesday, December 10, 2002 12:30 PM
To: Histonet
Subject: Dissolving skeletal muscle from bone

Hi all,

Does anyone know what solution to use to dissolve skeletal
muscle off of bone, so that the bone remains in tact.  This
is not for a histology study, but I thought that someone out
there on the histonet would know how to do this!

Thanks in advance,
Kim Merriam
TKT
Cambridge, MA

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Geoff
McAuliffe
Sent: Thursday, March 18, 2004 6:10 PM
To: Karl Garsha
Cc: histonet <@t> lists.utsouthwestern.edu; Microscopy <@t> MSA.Microscopy.Com
Subject: Re: [Histonet] digesting soft tissue

Household bleach, 1 part bleach to 4 parts water.
or
Adolph's meat tenderizer is papain, don't know the concentration needed.

You can get papain form Sigma, works well at 37C. I don't know any other

details.

Karl Garsha wrote:

> Greetings,
> I'm trying to find a good cocktail for non-specific digestion of
> tissue from bone samples--only the mineralized bone needs to be left
> unscathed.  I'm looking for a relatively cheap solution that will
> yield nice clean bones.  I'm aware of a product from Fisher, but it
> costs about $70/100ml. It seems to me that there must be a simple
> solution that I'm just not aware of (aside from culturing maggots).
> Thanks in advance for any suggestions.
> Regards,
> Karl G.
>

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff <@t> umdnj.edu
**********************************************

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