Dear oh Dear
On behalf of myself as a member of the uncivilized
cytometrists I appologize for my comments in advance.
The fact that a laboratory receives samples leaves me with
the impression that these samples are processed, analyzed
and sent back with a diagnosis. If you are not sure how to
deal with the sample I think not to accept it looks very
civilized to me.
Buying an airplane does not give me the ability to fly it,
let alone to offer an airline service. I happen to run
cytometers and do know quite a bit about the technology.
When I play with blood samples I use log/log scatter
displays and scatter versus fluorescence, so I would
probably obtain nice clusters a lot of people have never
seen, but still would not want to undertake bone marrow
analysis as I lack the handling expertise. If it would pay
my mortgage I would be happy to start learning about it and
ask perhaps the most basic questions about the gating
strategy and which reagents to use and from what
manufacturer. Luckily there are guidelines about, people
have done it and training initiatives like the STEP exist,
and so far even the most basic questions have been answered
here on the internet.
Unless the fact of receiving a sample has been
misinterpreted and all that fuss is a big misunderstanding
about someone who wants to establish a method, the
underlying problem is more an ethical than a technical
question. First, it is quite difficult to take negative
feedback, and therefore perhaps better given in private.
Secondly we should be far more aware of our own limitations.
I think in being frank to admit mine before undertaking a
task I gained more credibility than if I would not have done so.
Gerhard Nebe-v.Caron
Unilever Research, Colworth,
Sharnbrook, Bedfordshire
GB - MK44 1LQ
Tel: +44(0)1234-222066
FAX: +44(0)1234-222344
gerhard.nebe-von-caron@unilever.com
______________________________ Reply Separator _________________________________
Subject: Re: CLINICAL question: bone marrow
Author: bsherbur@west.bidmc.harvard.edu at INTERNET
Date: 20/05/97 02:00
Dear Adrian,
First of all, on behalf of the entire world of civilized cytometrists and
hematopathologists, we apologize summarily for the incredibly rude response
given by the individual in Seattle. I would be afraid to ask a legitimate
question if I knew that my practice would be inappropriately insulted. While
such acts of "roundsmanship" are unfortunately commonplace in academic medicine,
they are a detriment to the scientific process and a danger to patient care, as
everyone ends up refusing to admit when they don't know something!
Secondly, try gating on the lymphocytes and any population of large
mononuclear cells. Usually clinicians are looking for involvement by lymphoma,
or blasts in the case of leukemia or transforming myelodysplasia. We use FITC
anti-CD45 and a combo of PE anti-glycophorin and PE anti-CD14 to check our gate,
the PE labeled antibodies are to help ascertain when we are merely looking at
normal monos or nucleated RBCs. If you find cells in the lymph or small
mononuclear cell gate that have an order of magnitude dimmer LCA (CD45), there
is a good chance that they may be abnormal. If there's no history available
(and even if there is), we usually make a smear of whatever we get to see what's
in there. Another way to go if you do 3-color work is to gate on dim LCA
positive cells with low light scatter, i.e., to the dim LCA side of the lymphs.
This will allow you to pick up small numbers of blasts, if you're looking for
minimal residual disease. Otherwise, with no history, we look for a B-cell
clone or abnormal antigenic expression in the lymphocyte gate, as flow is more
sensitive than morphology for picking up low levels of lymphoma involvement.
Third, one of my favorite references for these issues is the now 2nd edition
of Flow Cytometry and Clinical Diagnosis by Keren, Hanson and Hurtubise,
available from the American Society of Clinical Pathology Press. You can call
and order one from them toll free, 800-621-4142
Best wishes,
Brad Sherburne
Co-Director of Hematopathology
Beth Israel Deaconess Med Center
Boston, MA
_______________________________________________________________________________
Subject: CLINICAL question: bone marrow
From: Adrian O Vladutiu <vladutiu@acsu.Buffalo.EDU> at smtplink-nedh
Date: 5/15/97 11:47 PM
We receive more and more bone marrow aspirates for flow cytometry
analysis. How does the group feel about the value of bone marrow analysis?
How does one gate cells (or analyze ungated cells)? In what conditions is
the bone marrow analysis most useful? Finally, are there references of
general interest regarding clinical value of bone marrow analysis by flow
cytometry?
Thank you in advance for your help.
Adrian Vladutiu
The Buffalo General Hospital
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:46 EST