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. 2012 Jul 25;13(8):9400–9418. doi: 10.3390/ijms13089400

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© 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

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Schematics of the Pulsed Interleaved Excitation-Fluorescence Lifetime Correlation Spectroscopy (PIE-FLCS) measurement. (A) Protocol for the PIE-FLCS measurements. (B) Example of fluorescence traces in the AF488 and AF647 channels. (C) TTTR methodology allows independent reconstruction and selection of fluorescence decay traces of AF488- (detector 1) and AF647-labeled (detector 2) proteins. (D) Weighting filters for correlation. (E) Lifetime-filtered autocorrelation curves in both channels.