Hello connection
While performing potency test by using BD FACS instrument some time we few population inside the gate few outside of gate and some time I got population outside of the gate I am not able understand why it happened like I attached one picture please refer that one
Please provide valuable suggestions
Thank you in advance
While previously used gates can serve as a good starting point, you will need to adjust them using your control samples for each flow cytometry experiment, as there is a certain amount of drift within the measurements.
I totally agree with Dr. John. I think you need to start with your control tubes and adjust your experiment setting based on it and gate your population then move to the sample tubes and collect the sample events. Also, you can discuss it in detail with the FACS team in your core facility may will set with you during the experiment and help you directly to gate your correct population. Finally, don't worry and this is normally happening during the research journey. Good Luck.
You can simply adjust your P1 gate in the first plot. If this is an analysis of cultured cells consisting of a single cell type you have possibly apoptotic/dying cells with low FSC & high SSC towards the left of gated population. If the sample contains different cell populations you can use the P1 gate to select your population of interest if you are able to identify it by the scatter properties. Yes, the use of proper controls would help you gate the correct population.
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