I need a little more information if I am to help with antibodies. What patient population will you be studying and how much clinical information will you get? Will you only have blood and bone marrow? This information determines the size of your panel. If you know that you are getting a chronic process you don't need to study a lot of antigens useful in acute leukemia and vice versa. The kits are expensive and not optimal for every case and you need to do at least 3 color panels. The important point is not just the antibody panel. You also have to have excellent standardized technique and know how to analyze the data to arrive at a correct clinical diagnosis (this is the most difficult). If you are doing bone marrows you might do the standard 45 gating, although other methods work well too. You have many questions (and t is good to ask them at the start) and I think you may need more than just a little advice. Where are you located? I think you could also use the names of good flow people near you to help out. It is very helpful to visit a good clinical lab and actually see how it is done. As for the questions that can be answered with no data- you can flow anything that has tumor in it. If there is tumor in the blood you can study it there. You should never ficoll out the mononuclear cells- use a whole blood lysis technique. You need to study the U.S. and Canadian Consensus Recommendations on the Immunophenotypic Analysis of Hematologic Neoplasia by Flow Cytometry, Communications in Clinical Cytometry 30:213(1997). This covers all the basics like whether or not you ficoll or what controls are recommended. It is a good place to start. I also recommend going to courses on clinical flow and attending meetings. Maryalice Stetler-Stevenson, M.D., Ph.D. Chief, Flow Cytometry Unit Laboratory of Pathology, NCI, NIH Sometimes you're the windshield, sometimes you're the bug. > ---------- > From: Mohammad Poureslami > Reply To: poureslam@yahoo.com > Sent: Thursday, December 13, 2001 6:10 PM > To: Cytometry Mailing List > Subject: Need your help > > > 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 > > > __________________________________________________ > Do You Yahoo!? > Check out Yahoo! Shopping and Yahoo! Auctions for all of > your unique holiday gifts! Buy at http://shopping.yahoo.com > or bid at http://auctions.yahoo.com > >
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