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Extra info for Flow Cytometry in Hematopathology: A Visual Approach to Data Analysis and Interpretation
At the same time, staining for the appropriate cytoplasmic lymphoid marker, either cCD22 or cCD3, should also be performed. The frequency of cases where additional testing for MPO Ab and cCD3 or cCD22 is needed is relatively low. Cytoplasmic CD3, CD22, or mu chain: With the use of multiple antibodies in the panel and the multicolor approach, the need to stain for cCD3 and cCD22 rarely arises in our laboratory. In most cases of T-ALL, the presence of CD2, CD5, and CD7 is sufficient to infer the T-cell lineage.
A,b) First set of antibodies: The intensity of the positive kappa light chain on CLL cells is inappropriately bright. The peak fluorescence of kappa is one decalog brighter than that of lambda (c). , dim positive kappa on CLL cells). The same results are seen on the corresponding kappa/lambda overlay histogram (d). 5 (a) The conjugation of CD10 to PE results in an apparent increase in CD10 brightness on the benign germinal center cells (lymph node with FRFH). The resulting CD10/CD20 staining pattern becomes similar to, rather than distinctive from, that seen in a lymph node with FCC lymphoma (b).
With some brands of kappa/lambda reagents, it may not be possible to achieve the appropriate fluorescence intensity. 4) and may fall in the same fluorescence range observed in FCC and MCL. The intensity of surface light chain expression is one of several criteria critical in the diagnosis and subclassification of mature B-cell malignancies. Therefore, in selecting the optimal polyclonal kappa and lambda antibodies, the appropriate fluorescence signal is a more important consideration than the ability to detect extremely dim light chain expression.