Dear Dr. Neumann If you have not already done so, a good start is to make sure there are no fluorescent solutions (like PH indicators) in the cell preparation. I recently had an experienced person bring me some cells that had been incubated and suspended in medium containing phenol red, and we could not get any usable reading. Joseph. At 06:34 PM 31/10/97 -0200, Jorge Neumann wrote: > We are testing some monoclonals developed in an affiliated >lab against myelogenous leukemia. We are trying to see if they are >specific against a particular sub-classification of AML (FAB M0 to M7), >labeling the cells with the monoclonal followed by the appropriate >conjugate plus a set of commercial monoclonals (CD7, CD13, CD14, CD19 and >CD34). Our problem is the high background fluorescence the cells are >presenting on incubation with the isotype controls, which are making the >interpretation of the results quite difficult. Is there any other >suggestion besides pre-incubating the cells with media plus FCS?. Any tips >are welcome. Thanks in advance. -- Joseph Webster Flow Cytometry Facility Centenary Institute
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