How to clean up cell fragments before cell sorting - (May/20/2015 )

Hello,

We are using Worthington papain dissociation kit to isolate the neural cells from adult mice (4wk - 8wk). Somehow, we always have a lot of cell fragments with the detached neurons in the final fluid. Based on DAPI staining, we knew that our procedure was not massively killing the cells. The person doing FACS for us said that all these cell fragments were also recognized by the FACS machine and about 80-90% detected events during the sorting were just junky pieces, which largely elongated the sorting time and hurt the quality of collected neurons. I am wondering how people handle this type of problem. Do you think a Percoll or Ficoll gradient centrifugation is recommended to be used to purify the neurons before FACS? Any other suggestions?

Thanks a lot!

Sincerely,

S

When you use FACS, the size of fragments and neurons are different. You could try which part you need to analysis.