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Fig. 5 | BMC Cell Biology

Fig. 5

From: Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)

Fig. 5

A. pallida cells labeled with fluorescent markers. The A. pallida cell suspension was incubated with the same fluorescent markers that were used to label coral cells and analyzed on FACS AccuriC6. All the samples were analyzed on two dimensional plots of the 488 nm channel (533/30BP) which detects the fluorescent markers and on the Far-red 640 nm channel  (675/25BP) which differentiates the Symbiodinium positive cells. Marked in the gates are the differentiated populations. The analysis was done after differentiation of cells and debris on FSC and SSC, and live/dead cells using PI. a) Background control of unstained cells in the gate are the Symbiodinium cells positive in the 640 nm channel (675/25BP) due to natural fluorescence. b) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells with the lowest level of metal was found to differentiate from the main population, labeled in the gate. c) Labeled cells with ALDEFLUOR a marker for ALDH activity. In the gate are low, moderate and high labeling populations. d) Cells labeled with CellRox, a label for ROS. In this case negative and positive populations are detected (positive marked). e) Cells incubated with glucose analog 2-NBDG. The cells are positive, but no clear differentiation. f) Cells co-incubated for 3 h with green fluorescent beads for analysis of phagocytosis. Cells positive for bead phagocytosis are gated

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