I am analyzing platelet activation markers CD62P and PAC-1 in whole blood using flow cytometry. Presently I am using calibration beads for the fluorescence. But I see some papers reference the use of beads for instrument standardization. Are these beads essential? What things do I need to look out for to achieve quality, reproducible analysis?
Hi, why you want to use the calibration beads? for population gating? or ? I think you can stain platelet specific marker such as CD41 or CD61 at the same time, that may help you to identify the platelet population. In addition, you can use thrombin to as a CD62p positive control.
The beads used for instrument standardisation are needed if you are using two or more fluorochromes in the same tube. The calibration beads check for and minimise overlap in the emission spectra of the chosen markers. This is for all flow work, not just platelet flow work.
I am using CD61 for platelets identification and I don't have an problems with setting my gates. I understand I need to run beads with my samples to ensure the instrument setting has not varied over time as I will be running my analysis over several weeks. I was wondering if there are other ways of monitoring the instrument setting. If not, what beads do I need as there appear to be different types?
I just know beads to be used to calibrate the facs aps for different channels in case you don't want to use/ you don't have appropiate IgG controls
regards
Sven
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