How to set the gate for rathippocampal neuron apoptosis assay?

Annexin V assay is normally used for cell line samples, thus only debris need to be excluded by gating. Since there aremultiple types of cells in tissue samples, it depends on the purpose of experiment, whether it means to detect apoptosis of the whole tissue sample or some specific cells. Cells can be gated by FSC/SSC or some other biomarkers. Besides, operation of tissue sample preparation may also effect the result, Inappropriate operation may also result in false positives due to sample handling.