Protocol Online logo
Top : New Forum Archives (2009-): : Flow Cytometry

DNase and PMA/Iono - (Oct/18/2009 )

Hi all,
I've read that adding DNase I during or after PMA/Iono stimulation can avoid the formation of clumps, thus improving flow cytometry analysis. Do you have experience with this protocol? Could you explain this to me in more details?

Thanks a lot!


I haven't heard that one before but I can offer an educated guess....
Dying cells release DNA which is very sticky and will cause cells to stick together (in snot balls)
PMA and iono treatment can be pretty toxic to cells (at least it has always been to T cells when I do intracellular cytokine stains) and cause a lot of death and DNA release.
DNAse treatment will clean up the sticky DNA

DNAse treatment is pretty common in a lot of single cell isolation protocols b/c of the cell death/sticky DNA/ snot ball issues.