Need your help

From: Mohammad Poureslami (poureslam@yahoo.com)
Date: Thu Dec 13 2001 - 18:10:05 EST


Dear colleagues,
I'm working In a university teaching hospital, at
hematology&oncology ward. We have recently equipped
with a bench top flowcytometer(one argone laser) to
work mostly on patients with leukemia. I'm fairly
familiar with flowcytometry, however I have no
experience working on leukemic samples. We have
purchased Acute luekemia phenotyping kit from Becton
Dickinson company, in which monoclonal antibodies such
as CD3,CD19,CD22,CD20,CD5,CD7,CD10,CD13,CD33,HLA-DR,
and two vials of negative controls are included. Half
of these reagents are conjugated with FITC, and the
other half with PE. I'm trying to set up a protocol
for acute and chronic leukemia, but I need your help.
I know that this kit is not appropriate for chronic
leukemia, and I am also not sure the kit that we have
now has a good combination for acute leukemia
phenotyping. So in this regard, I would like to know
the latest antibody combinations for those
phenotypings. We can use up to three
colors(FITC,PE,PerCP)on our system. I also need to
know if I have to work only on BM samples of these
patients, or if I can also use their PB cells. Do I
have to separate their mononuclear cells? Do I need to
run a normal sample every time I run my patients'
samples? and if the answer is yes, what good can do
this normal sample? I'm assuming that none of the
gatings drown on this sample would be used on patient
sample. So as you see I have many questions that I'd
like to get their answers. I don't know whether it's
possible to get a detailed working procedure on those
applications that I mentioned above. A protocol that
detailes everything from the very beggining to the
very end.
I'm anxiously looking forward to hear from
you and if you don't mind keep in touch with you when
I need your consultation. Your valuable time and
consideration are highly appreciated.
Best Regards,
F. Osati


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