Dear All, Has anyone tried using "Accumax" from TCS Cellworks at all in flow cytometry? It is supposed to be an anticlumping reagent, recommended for using for cell counting, which requires little or no Ca++ or Mg++. The cells are not suitable for culturing after disaggregation in this reagent. Anyone? My little bit on clumping on the sorting front; We sort cultured marrow cells which can be horribly clumpy and sticky (hence my wondering about Accumax) and we sort with a suspension at 10x10e6cells/ml having filtered the cells immediately before putting on the machine and the proportion of dead cells definitely affects the amount of clumpiness which develops. We don't put on more than 2ml at a time, and during a run take off the tube and give it a very gentle mix if it is really settling out, rather than using the buzzer on the machine. They can always get a second pass through a filter. The on-line thing sounds like a big faff if you get it clogged up so we've never done it. Since persuading everyone of the need to filter, before running each and every sample, we haven't had ANY blockages. (Famous last words). Carolyn Jefferiss Carolyn Jefferiss Ph. D. Pharmacy and Pharmacology University of Bath Claverton Down Bath BA2 7HY
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