[Histonet] Re: bone marrow aspirate

[Bob Richmond]

I don't do the procedure any more, but earlier in my career I did
about 400 iliac crest aspirations and biopsies, reading them all
myself. I always had an assistant, whom I often trained myself - you
really can't both tend to the patient and make the smears. If the
assistant didn't have her own method, I'd teach her to squirt some of
the aspirate onto a slanted slide, pick up the particles with a broken
applicator stick and move them to additional slides to make smears.

I'd then squirt the rest of the still unclotted aspirate into neutral
buffered formalin, which of course doesn't clot blood, so that you get
a suspension of tiny particles in clear liquid. This could be poured
into a tea bag (or nowadays into one of those little nylon bags) for
further fixation and subsequent processing.

In those days I'd also put the bone biopsy specimen and a clot into
Zenker/Helly fixative, which of course is no more all gone. I don't
see any reason to process clots today, unless the aspirate clots
prematurely.

Bob Richmond
Samurai Pathologist
Knoxville TN
***********************************************
> Message: 4
> Date: Fri, 4 May 2012 16:24:05 -0500
> From: "Troutman, Kenneth A" <Ashley.Troutman <@t> Vanderbilt.Edu>
> Subject: [Histonet] bone marrow aspirate
> To: "Histonet <@t> lists.utsouthwestern.edu"
>        <Histonet <@t> lists.utsouthwestern.edu>
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>        <7B310892042DA74CB3590053F424CFE6145CB28982 <@t> ITS-HCWNEM06.ds.Vanderbilt.edu>
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> Hi Tim,
>
> The best way I have found over the years actually requires the person collecting the specimen to do the most work.  What we used to do is after the aspirate is performed, make all of the smears, and then inject the remaining aspirate directly into formalin before it coagulates.  This gets rid of all the blood and ensures all that is left is marrow.  After sufficient time in formalin, filter the marrow out of the formalin and process.
>
> As for a processing protocol, we do a run as follows:
> Formalin 30 min
> 70% Alcohol 20 min
> 90% Alcohol 10 min
> 100% Alcohol 10 min
> 100% Alcohol 10 min
> 100% Alcohol 15 min
> Xylene1 15 min
> Xylene2 15 min
> Xylene3 20 min
> Paraffin1 15 min
> Paraffin2 15 min
> Paraffin3 30 min
>
> This protocol was done with pressure/vacuum.
>
> We have excellent results with this and the pathologists do not have to spend a lot of time hunting for small areas of marrow on the slide, the whole slide is marrow.
>
> Good luck!
>
> Ashley
>
>
> Message: 5
> Date: Fri, 4 May 2012 16:54:57 +0000
> From: "Coskran, Timothy M" <timothy.m.coskran <@t> pfizer.com<mailto:timothy.m.coskran <@t> pfizer.com>>
> Subject: [Histonet] bone marrow aspirate
> To: "histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>"
>                <histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>>
> Message-ID:
>                <70249E5B79AFEB48A47D78568CE216E9027FB1 <@t> NDHAMREXDE02.amer.pfizer.com<mailto:70249E5B79AFEB48A47D78568CE216E9027FB1 <@t> NDHAMREXDE02.amer.pfizer.com>>
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> Does anyone have a protocol on how to fix and process a bone marrow aspirate to paraffin?
>
> Thanks,
> Tim Coskran
> Pfizer