Fluorescence-Based Yeast Cell Analysis

Fluorescence measurements were taken using a Becton Dickinson (BD) special order cytometer with a 514 nm laser exciting fluorescence that is cut off at 525 nm prior to photomultiplier tube collection (BD, Franklin Lakes, NJ). Events were annotated, subset to singlet yeast using the FlowTime R package (Wright et al., 2019 ). A total of 10,000–20,000 events above a 400,000 FSC-H threshold (to exclude debris) were collected for each sample and data exported as FCS 3.0 files for processing using the flowCore R software package and custom R scripts (Supplementary file 1; Havens et al., 2012 (link); Pierre-Jerome et al., 2017 (link)). Data from at least two independent replicates were combined and plotted in R (ggplots2).