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Jun Ouyang
11-16-1998, 03:21 PM
Dear colleagues,

Thank you for all your kindness responses to my question on "how to
clean the bone marrow?". And I just summary the methods provided to me.

Thank you again.

Jun Ouyang
************************************************** **************
I wrote:

Hello colleague,

I am research fellow at University of Hong Kong now. I am doing some
research work on the relationship between the fatigue and the
trabecular bone on human lumbar vertebrae. But when I make the
trabecular bone sections, I could not remove the bone marrow and other
soft tissue on the trabecule. I have tried the ultrosonic method and use
2% hydrogen peroxide at 37 centigrade degree for 24 hours. But it still
looks very "dirty" under the microscope.

So I would appreciate any help on this problem from you. Thank you very
much.

Jun Ouyang MD & PhD
> Medical Biomechnics Lab
> Department of Anatomy
> The First Military Medical University
> Guangzhou, 510515
> P.R,China
> E-mail: jun_ouyang@163.net


Followings are response:
************************************************** ******************
Hello Jun

I am PhD.student in biomechanics at EPFL in Lausanne/Switzerland. I
succeeded to remove marrow in trabecular bone
by using (until 4 times) the following procedure:

put the bone samples in a soapy water solution (use liquid soap for the
hands that is in use in restrooms) and put
everything in the ultrasound bath (directly, not in a glass can !!!).
apply the ultrasound for about 15 min at 30 degrees.

remove the soapy water solution from the ultrasound bath and replace it
by 1:10 mixture with 5%-"Eau de Javel" and
water ("eau de Javel is also called "hypochloride bleach", here in
switzerland it can be bought in a pharmacy and is very
cheap). apply the ultrasound bath again for 6 minutes.

repeat this procedure several times. normally you should see an
improvement. but profit the maximum intensity of the
ultrasound wave by NOT putting the solutions and the samples in a glass
can, and then putting the glass can in the
ultrasound bath !!!!

best regards and good luck Stefan

************************************************** ********************

Hi Dr. Ouyang,

Try to use bleach, such as the stuff you would buy to use on clothes. It
has
been used by our lab to clean soft tissue (such as cartilage, which you
should score/roughen up to allow the bleach to reach different layers).
Just
leave the bone sitting in it at room temperature for 24-36 hours. Leaves
the
bone clean. Another method (used by the Zoology Department here) is
placing
bones in a "bug box" where the box is filled with some type of bugs.
After a
week the bones are sufficiently clean, and after washing with simple
soap
and water (to get the bug feces of the bone), they are ready to be set
out
on display. If you want more info on this method I can probably track
down
an email address for you to contact someone in that department.

Good luck,

Dan Magnusson B.Sc.
Master's student in Biomechanics
Human Performance Laboratory
Faculty of Kinesiology
University of Calgary
Calgary, Alberta, CANADA
T2N 1N4
Tel: 1 403 220-3432
Fax: 1 403 284-3553
email: dan@kin.ucalgary.ca
************************************************** ******************
Jun

You asked Biomch-L about methods of cleaning bone before testing.

The following paper may be of some help:

D J Sharp, Tanner and Bonfield
Measurement of the density of trabecular bone
J Biomechanics
Vol 23, No 8, pp853-7, 1990

This paper was concerned with methods of defatting for density
measurements
of trabecular bone, and found that trichlorethylene in an ultrasound
bath
for 4 hours removed all the fat (1 hour trichlorethylene in US bath
didn't,
nor did 4 hours of alcohol in US bath. Samples 9.5mm dia., 5-10 mm
long).

Is it necessary for the bone to look or even to be clean? Incidentally,
as
I'll shortly be doing some work with trabecular bone: What are your
section
thicknesses and how are you testing them? Hope that this is some help.
Do
let me know, and please post a summary of responses to the list.

AJ Stephenson
aj.stephenson@btinternet.com
Department of Plastic Surgery
Northern General Hospital NHS Trust
Sheffield
UK
Phone: +44 (0)114 2434343 pager 751
************************************************** ***************
We have had good success with a 50% solution of household bleach
and water. This is particularly effective when supplemented with
ultrasonic agitation. You must check this every 30 min or so since
the procedure will eventually completely destroy the bone.
Good luck

John A. Hipp, PhD
Baylor College of Medicine
Houston, TX USA
************************************************** *********************
As for defatting the bone, we generally use ethyl ether. I don't
know if that would work for your application, but you might give
it a try.

Keep in touch.

Dale

C. R. 'Dale' Bass
Senior Scientist
Automobile Safety Laboratory
University of Virginia
Phone (804) 296-7288
Fax (804) 296-3453
e-mail:crb7q@node3.unix.virginia.edu


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