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  • summary of clean bone marrow.

    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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