Cell Counting Using Flow Cytometry

Absolute microalgae cell counts were obtained for WT and LOG OX1 cultures at 14 dpi and stationary phase for three biological replicates each by a method described in an earlier study (Chioccioli et al., 2014). Absolute microalgae cell counts were obtained by adding an internal microsphere cell counting standard (Count Bright; Invitrogen, CA) to the flow cytometric sample (single platform testing). A 1:9 dilution of Count Bright suspension was added to the microalgae culture. Using a FSC-A vs. SSC-A plot to separately gate cell events vs. bead events, the ratio of bead events to cell events (together with the known concentration of beads) was used to calculate the absolute cell concentration. This experiment was repeated three times.

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