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We do a lot of cell suspension preparation from lymph nodes and usually do not
get clumping. However, when we first started we did and discovered that we were
trying too hard to disaggregate the tissue. We began to be more gentle in our
method and that solved the problem. We gently press the tissue through a metal
sieve. Other labs cut up the node with two scalpels using a scissors action or
simply put small pieces of node in a culture tube with media and shake. These
will all give a cell suspension. You will need to filter out connective tissue
or allow it to settle. Many labs use 50-70 micron nylon filter material for
removing clumps. You can obtain the filter material from Tetko in Depew, NY,
(716) 683-4050 or from Small Parts in Miami, FL, (305) 751-0856 and you can
obtain ready made, disposable funnels with filter material in them from Dako
called filcons, (805) 566-6655.
Hope this helps,
Tony Bakke, PhD
Clinical Immunology and Flow Cytometry Lab
Oregon Health Sciences University
bakkea@ohsu.edu
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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