Joy Mundy asked, "Can anybody help me with a cell suspension solution that will inhibit cell clumping." ---------------------------- 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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