Fig. 4
From: Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)
12 coral cell populations sorted by FACS. P. damicornis cell suspension was labeled with DAPI, Cellrox, and LysoTracker Deep Red, analyzed and sorted on FACS ARIA II. Sorted cells were put on ibidi 18 microwell plate coated with poly-L-lysine, overnight at 4oC prior to obtaining the cell images by light microscopy (image inserts). a) Light microscopy image of unsorted cells. b) After excluding cell debris by their intrinsic size (FSC) and granularity (SSC), we gated a two dimensional plot of the 633 nm (755LP 780/60BP) channel containing the Symbiodinium positive cells and DAPI stained cells in the 405 nm (450/50BP) channel. We sorted 3 Symbiodinium positive populations (upper panels, p6–8), and two positive populations for DAPI (lower inserts, p3 and p1). c) Ungated cells in panel b were analyzed on two dimensional plot of 488 nm (505LP 530/30BP) channel- ROS and 633 nm (660/20BP) channel- LysoTracker. An additional 6 populations were differentiated by this panel (p4–5 and p10–13). d) Population 12 (P12) was additionally separated into two populations on panel of size (FSC) and LysoTracker (APC) to P14 and P15. Notably, P14 is enriched for nematocyst cells. Red scale bar for all inserts = 100 μm