Hi flow collegues, Our pathologists have decided to "rekindle" flow cytometric analysis on bladder washouts after they met with the urologists in this area. Apparently there was a fairly recent consensus review regarding the use of bladder washouts as a surveillance tool. The plan is to do ploidy analysis on: 1. initial bladder biopsies with grade I or II tumors 2. bladder washouts for surveillance My questions are: 1. Which procedure is the most likely to avoid false "near-diploid/aneuploid" populations as produced sometimes by the differential PI staining of large squamous epithelial cells? I have read a very good article by a german group (Liedl et al, Urol Int 1995;54:22-47) who was using a PI - Cytokeratin 8,18 (indirect?!) dual staining procedure. Apparently this procedure excludes the squamous cells in addition to the inflammatory cells. I personally like the dual staining idea but have had some problems years ago with the compensation adjustment. 2. Which cytokeratin antibody is the most popular one to use for bladder washouts. Is it really the CK 8,18 and is it available directly conjugated (the paper descibed it indirect)? We have Dako's MNF 116 on hand, can we use this? 3. How does one interpret near-diploid populations? I feel very comfortable in calling near-diploid populations aneuploid in solid tumors and other neoplasms since we usually get excellent C.V.s. But I have to admit that I get very nervous about interpretations of ploidy patterns in bladder washes since we had so many problems years ago when we were still doing them. Wide C.V.s, differential PI staining of the various subpopulations, "crappy" histograms etc. I would really appreciate you taking the time to answer my questions. I realize that I could have read a lot more references than the ones I have researched myself as well as try different procedures and see how each one works. I hope nobody feels that we clinical labs just want a quick answer without doing the "dirty work". But it really is a matter of time we often just don't have. So, on behalf of all the clinical labs who feel the same way, I would like to thank all of the participants providing this wonderful resource! Thanks - Andrea Andrea Illingworth Dahl-Chase Diagnostic Services/Flow Cytometry 333 State Street Bangor, Maine 04401 (207)990-4855
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