Figure 2.
Flow Cytometry Gating Workflow for Microbial Particle Enumeration. a) An unstained aliquot of the dialysis effluent specimen is measured, and a SYTO® 9 positive gate is set based on the upper limit of the unstained 488nm Ex 530/30 Em fluorescence. Sample is then backgated to ensure that no significant auto-fluorescent population remains. b) The SYTO® 9 positive gate is inherited to the data from a stained aliquot of the dialysis effluent. The specimen is backgated for demonstrative purposes (right panel). c) A bivariate plot (FSC-A vs FSC-H) is used to exclude doublets (left panel), unstained data is used to determine the threshold for Live/DEAD Violet staining (middle) and double positive gating applied to allow for determination of viability status (right).