Hi Bill
We've had the most luck simply placing the tissue in a petri dish with a
little RPMI and gently tearing it apart with two (preferably toothed) forceps.
This works well for nodes, spleens and all kinds of tissues. If it is too
bloody, i.e. splenic tissue, we incorporate a red cell lysing step. Viability
is good, including the large cells, which are usually the elements of interest.
With tissue grinders and mesh, we always ended up with just the small reactive
T-cells in B-lineage large cell lymphomas, and had to resort to immunoperoxidase
for typing. But with mechanical dissociation, we usually can find some clonal
B-cells or aberrant T-cells corresponding to the lymphoma, and can back-gate to
prove that the population of interest contains the larger cells by light
scatter. One caveat is that we like to sample from opposite a face which is
sampled for histology to make sure that lesional tissue is present, and always
correlate the two. We still need immunohistochemistry on about one third of our
large cell lymphomas and all of our Hodgkins disease cases, because they are too
fibrotic and the cells of interest just don't survive the process. I too would
love to hear of anyone's favorite processing technique!
Brad Sherburne, Beth Israel Decaoness
Medical Center, Boston
bsherbur@west.bidmc.harvard.edu
_______________________________________________________________________________
Subject: Lymph Node Dissociation Methods
From: Bill Justice <bjustice@kumc.edu> at smtplink-nedh
Date: 7/29/97 06:35 AM
Hello,
I am asking this question on behalf of another flow cytometry operator at our
institution.
This is a very basic question. When processing lymph node for L/L panels, what
are other labs doing to mechanically process the node?
Tissue Homogenizers?
scissors and scalpels?
pushing through wire mesh?
What?
What are the pitfall and potential problems with difference techniques?
Thanks.
Bill Justice
Flow Cytometry/PC Support
KUMC Clinical Labs
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