What concentration does the PI have?
We have BD FACSCalibur and Muse Cell Analyzer(Millipore). We have used Muse cell cycle analysis kit to Muse Cell Analyzer. The kit includes PI. So I tried the kit for BD FACSCalibur. After staining, I washed the reagent with PBS, and meshed it. Exactly, I should purchase normalized kit for BD. Could I use the kit to BD?
Hello Yusuke, Muse uses a green emission laser to excite fluorochromes. Some of them, like PI, can be excited by 488 nm laser like Fascalibur has.The problem with Muse kits is that they are adapted to work with few cells, and maybe, in your case, as you want to run in a FACScalibur you are using more cells than kit can stain. PI staining for cell cycle must be stequiometric, therefore you must be assure that there is enough PI for all cells. Hope it helps, Alberto
Dear Dr.Alberto, Thank you very much for your answer. For FACScalibur, I will use more cells than used to be done in Muse analysis. Equilibrium PI staining will be done.
Hi Yusuke, why not? you can detect PI in FL!, FL2, and FL3 using the good old Calibur. If you use FL2 in lin scale and FL3 log you are able to gain information about cell cycle AND apoptosis (sub G1 peak). As Alberto was mentioning you have to watch that you have enough PI. PS: Be good to your flow cell and clean not only with 10% bleach for 10min, but also with 70% Ethanol to prevent build-up.
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