I have also been surprised by unremarkable flow data from an FNA of an obvious large cell lymphoma (phenotyped as B cell by immunocytochemistry). I like the idea that the cells were fragile. No population of large cells by forward scatter were apparent. /Michael Kornstein, MD, Medical College of Virginia, Richmond On Wed, 8 Oct 1997, Brent Dorsett wrote: > > > >Viki Mosiman > >Robert H Lurie Cancer Center > >Flow Cytometry Core Facility > >Northwestern University Medical School > >Chicago, IL > >vlm646@nwu.edu > > > >I am interested in an opinion on the best way to detect Large-cell > >lymphoma(B Cell type) in lymph nodes. > > > >We currently disaggregate nodes then surface stain using > >45/19/kappa/lambda. Although > >we often have success ,there are cases where clearly the node is a > >malignant B-cell lymphoma by morphology in Surgical Pathology but we see > >no evidence in Flow. We have tried blocking with fetal calf and mouse > >serum. We have tried different gating strategies such as 45ss combined > >with 19ss gating or combined with fs. We have tried different staining > >combinations -19/5/kappa/lambda-45/19/10/kappa&45/19/10/lambda. We have > >tried live dead assays(7AAD) combined with surface staining in case a node > >was necrotic. Someone even suggested injecting nodes with PBS to express > >cells instead of mincing and grinding lymph nodes(they thought this might > >be a more gentle process). > > > >We don't seem to have the same problem with other malignant lymphomas in > >lymph nodes,just with the diagnosis of Large-cell lymphoma ( B-Cell type). > >Any suggestions would be greatly appreciated. Thanks for your help! > > > >Clinical Flow Lab > >Northwestern Memorial Hospital > > > > > Viki, > > Recently we have seen several cases where the malignant population in large > cell lymphomas was extremely difficult to demonstrate in flow. One, a lymph > node, was easily demonstrated by immunohistology to consist of large numbers > of B cells infiltrated with reactive T cells. In flow cytometry after the > usual mechanical disaggregation, suspesion in RPMI 1640 + 5% FCS, filtering > through 100 micro nylon mesh and washing almost no malignant B cells could > be detected. The second was a FNA of a mass in the liver collected into RPMI > 1640. This too immunostained positively as large B-cells, but in flow was > composed of 99% small to medium T-cells. Cell viability was good in both > preparations but there appeared to be substantil very small debris. I'm > guessing that these large cells were particularly fragile -- although the > appearance of the FNA cells in histology seemed normal. I would be > interested in the experience of other with large cell lymphomas. > > Brent Dorsett > Special Pathology Laboratory > Lenox Hill Hospital > NYC > >
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