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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web