Re: BONE MARROW PROCESSING

From: Wing-keung LEE (hkleewk@netvigator.com)
Date: Wed May 05 1999 - 13:45:16 EST


Hello, Rachel:
I don't know whether this is the best way of bone marrow specimen
processing, but want to share my experience.
1. Wash the specimen twice with pH7.4 PBS. I use the Immunfuge to spin 2
min at low speed after washing. Ensure no significant cell loss during
these steps. You can countercheck the cell population by prepareing a
blood smear. Also alert that high centrifugal force will casue decrease
in Forward scatter. Suspend the cell with protein base buffer e.g. fetal
calf serum or cheaper use 10% IgG free albumin in PBS.
2. Filter is unnecessary as you use fine pipette tip to aspirate the
sample. It is the cheaper and easier method. I found there is no
blockage of flow path or loss of cell population. I'm afraid that cell
will loss during filtration.
Good luck.

Best Regards,
LEE
United Christian Hospital,
Hong Kong
 

Gema Acebedo Lerzundi wrote:
> 
> The question is about the Bone Marrow sample preparation.
> What is the best method for Bone Marrow sample preparation prior to
> antibody staining ?
> 1. wash the sample first?
> 2. filter through needle or milipore filter?
> 3. both ?
> 4. neither of both.
> 
> we shall be grateful for your indications.
> 
> Gema Acebedo and Carlos Palacio
> Hematology Department. Flow Cytometry Unit.
> Hospital Vall d'Hebrón. Barcelona (Spain).
> Fax: +34932746015
> E-Mail: gacebedo@hg.vhebron.es



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