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Flow Cytometry Applications

Profiling Starvation & Recovery

Cyto-ID™ Green signal increases ~3 fold after 2-hour starvation. Starved cells placed back in growth medium for 1 hour demonstrate complete recovery.

Treatment Mean Signal (FL1) n=3
Control 83±5
2 hr. starvation 204±8
2 hr. starvation
& 1 hr. recovery

Autophagic Activator Factor* : 59

* Common parameter used for Multi-drug Resistance by Flow Cytometry. See formula below:

Profiling Autophagic Flux

By combining an autophagy inducer (starvation) with an inhibitor of lysosomal degradation (chloroquine) the assay can easily distinguish between increases in autophagic flux vs. autophagic vacuole accumulation.
Chloroquine blocks the lysosomal degradation pathway and normal autophagic flux is disrupted, resulting in auto-phagosome accumulation.
The mean FL1 signal from starvation with chlororquine increases much higher than starvation alone, indicating enhanced accumulation of autophagosomes resulting from the increased flux.

Treatment Mean Signal (FL1) n=3
Control 81±2
2 hr. of 300 µM chloroquine 129±12
2 hr. starvation 223±7
2 hr. starvation
with 300µM chloroquine

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