Hi! I'm a newborn in the flow cytometry context and have a few questions I haven't been able to find answers to inspite of a massive litt search. The aim of my diploma work is to do apoptosis (with Annexin V-FITC and PI) and cell cycle analysis (with PI) on animal cells in batch culture using a partec flow cytometer without a sorter. Concerning cell cycle analysis: -After fixtion in EtOH the cells should be treated with RNase. Does the RNase have to be DNase free? -Is it necessary to buy unstained trout red blood cells (TRBC) to use as a control for the method/flow cytometer or are TRBC only used in clinical applications? -Would a certain channel number be recommended for cell cycle analysis (e.g 255,512,1023)? Concerning apoptosis studies: -To set up the flow cytometer and compensate for crosstalk I'm going to use apoptosis-induced cells stained with FITC only and apoptosis-induced cells stained with PI only and run them separetly on the flow cytometer. I was recommended to use butyric acid (5mM) as an apoptosis inducer. Does anybody know an approximate incubation time to achieve an "apoptosis" maximum (i.e. where the signal will be the strongest)? Many thanks! Best regards, Ulrika Eriksson Division of Bioprocesses Institution of Biotechnology Royal Institute of Technology Stockholm, Sweden --
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:20 EST