Figure 3. Representative figures of gating strategy and identification of sperm cells during viability analysis using Live/Dead Viability Kit. In the dot plot (A) single cells were gated by plotting FSC-H (linear) vs. FSC-A (linear) to discriminate doublets. Further, single cells were gated for morphology (FSC-A linear vs. SSC-A logarithmic, dot plot B) separation of sperm cells (high SSC signal) from debris (low FSC and SSC signal). Last, we plotted CFDA or SYBR14 (FITC-A) vs. PI (PI-A) in dot plot C) to identify intact (FITC-A positive, PI negative) and dying/dead (FITC-A dim/negative, PI-A positive) sperm cells.
