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Obtaining quality cell suspension from fresh and/or frozen tissues is an important first step for subsequent 

analysis such as flow cytometry analysis (FACS) and nucleic acid purification. Cell suspensions can be

isolated from tissues by one or a combination of the following three mechanisms: chemical tissue

dissociation, enzymatic digestion and physical separation. Many methods are unduly tedious and time

consuming. Most importantly, cells in the tissue especially frozen tissue are usuallydamaged and show low

integrity and viability. It is very difficult to obtain high viability cell suspension from frozen tissues with high

connective tissue contents such as liver, kidney and brain tissues. We have developed a simple and rapid

method for isolating cell suspension from fresh or frozen tissues using a combination of chemical tissue

dissociation and physical separation mechanisms. The protocol can be finished in about 20 min with high

cell integrity and viability.

Morphologies of isolated cells from fresh/frozen mouse tissue using tissue dissociation reagent or 1 X PBS

with 2% BSA. The cells were stained with trypan blue.