Is there any evidence for these cells to be in the prep
based on light microscopy. I don't know if they would have
to appear bigger in light scatter peak or area signals as
that could also depend on detected scatter and the
refractive index of the cells.
Is there actually somewhere a systematic comparison on
lymphnode disaggregation methods? Unfortunately I still
haven't got easy CD-Rom access, but remembering the e-mail
summaries on the Purdue CD-Rom 3, is there a summary of the
tissue prep. issues? or has anybody a good literature
source?
Thanks in advance for any replies
Gerhard.Nebe-von-Caron@unilever.com
______________________________ Reply Separator _________________________________
Subject: Re[2]: large cell lymphoma
Author: mann0002@mc.duke.edu at INTERNET
Date: 09/10/97 22:36
We have had the identical experience and likewise have not seen evidence of a
large cell population by light scatter characteristics.
Karen
Karen Mann, MD PhD
Duke University Medical Center
Durham, NC
_______________________________________________________________________________
Subject: Re: large cell lymphoma
From: mkornste@hsc.vcu.edu at internet
Date: 10/09/97 6:50 AM
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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Subject: Re: large cell lymphoma
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