Dutch Boltz - Molecular Devices

Conclusions
The addition of a 409nm krypton laser onto a FLIPR-384 instrument has provided our
laboratory with the means to utilize FRET based voltage sensor probe technologies as a
screening tool for identifying ion channel modulators in an HTS format. As an initial study
we developed a membrane potential kinetics screen for the modified FLIPR that utilizes a
FRET-based coumarin-DiSBAC 2 dye system to screen inhibitors of ion channel X expressed
in HEK293 cells. Results from the study indicate that the percent inhibition of
depolarization for channel X compounds generated from data collected on the modified
FLIPR-384 instrument correlates well with that obtained from the standard kinetic plate
reader. The modified FLIPR, in addition, had comparable sensitivity and a 12-24 factor
increase in throughput. The ability to run the instrument in ratiometric mode in conjunction
to imaging all 384 wells simultaneously eliminates concerns such as variability and run down
of signal over time which was observed on the other plate reader during channel X
screening. These findings demonstrate the effectiveness of the modified FLIPR as an
alternative HTS platform for FRET based ion channel screening. In addition, we could use
virtually any dye on the modified FLIPR with the possibility of simultaneously following two
distinct physiological sensors.
References
Gonzalez, J.E. and Maher M.P. Receptor and Channels, 8: 283-295 (2002).
Mattheakis L., Savchenko, A. Current Opinions in Drug Discovery and Development, 4(1): 124-134 (2001).
Gonzalez, J.E., Oades K., Leychkis, Y., Harootunian, A., Negulescu, P.A. Drug Discovery Today, 4(9):
431-439 (1999).
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