Recombinant differential anchorage probes that tower over the spatial dimension of intracellular signals for high content screening and analysis
Anal. Chem.. 2009-12-01; 81(23): 9590-9598
DOI: 10.1021/ac9015227
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Recombinant fluorescent probes allow the detection of molecular events inside
living cells. Many of them exploit the intracellular space to provide positional
signals and, thus, require detection by single cell imaging. We describe here a
novel strategy based on probes capable of encoding the spatial dimension of
intracellular signals into “all-or-none” fluorescence intensity changes
(differential anchorage probes, DAPs). The resulting signals can be acquired in
single cells at high throughput by automated flow cytometry, (i) bypassing image
acquisition and analysis, (ii) providing a direct quantitative readout, and (iii)
allowing the exploration of large experimental series. We illustrate our purpose
with DAPs for Bax and the effector caspases 3 and 7, which are keys players in
apoptotic cell death, and show applications in basic research, high content
multiplexed library screening, compound characterization, and drug profiling.