I use height (FSC-H) plotted against FSC-A (area) to gate singlets, then analyzing PBMCs or T cells but I see there is the option of w for FSC and SSC. Is it useful?
The width (W) parameter can be use to gate or gate out the double or multiple cells from single cells. Based on that you can get more sensitive and precise results for flow cytometry studies. I use FSC-A (x-axis) plotted against FSC-W (y-axis) to separate single and double cells.
The width (W) parameter can be use to gate or gate out the double or multiple cells from single cells. Based on that you can get more sensitive and precise results for flow cytometry studies. I use FSC-A (x-axis) plotted against FSC-W (y-axis) to separate single and double cells.
The purpose of both FSC-H and FSC-W is to check the doublet or multiplet cells, based on FACS machine people generally use FSC-A vs FSS-W or FSC-A vs FSC-H to exclude multiplet. It is recommended that if FSC-A vs FSC-H is used then for better resolution of multiplet one should also use SSC-A vs SSC-W/H. For further information you can look BD flowcytometry guide.