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ZX Du, CQ Dong, JC Ren (2017)
A study of the dynamics of PTEN proteins in living cells using in vivo fluorescence correlation spectroscopyMeth App Fluoresc, 5
S Ramadurai, A Holt, V Krasnikov, G Bogaart, JA Killian, B Poolman (2009)
Lateral diffusion of membrane proteinsJ Am Chem Soc, 131
D Magde, E Elson, WW Webb (1972)
Thermodynamic fluctuations in a reacting system – measurement by fluorescence correlation spectroscopyPhys Rev Lett, 29
S Chiantia, J Ries, P Schwille (2009)
Fluorescence correlation spectroscopy in membrane structure elucidationBiochim Biophys Acta, 1788
T Krouglova, J Vercammen, Y Engelborghs (2004)
Correct diffusion coefficients of proteins in fluorescence correlation spectroscopy. Application to Tubulin Oligomers induced by Mg2+ and PaclitaxilBiophys J, 87
A Michelman-Ribeiro, D Mazza, T Rosales, TJ Stasevich, H Boukari, V Rishi, C Vinson, JR Knutson, JG McNally (2009)
Direct measurement of association and disassociation rates of DNA binding in live cells by fluorescence correlation spectroscopyBiophys J, 97
J Kubečka, F Uhlik, P Košovan (2016)
Mean squared displacement from fluorescence correlation spectroscopySoft Matter, 12
M Wahl, I Gregor, M Patting, J Enderlein (2003)
Fast calculation of fluorescence correlation data with asynchronous time-correlated single photon countingOpt Express, 11
V Dunsig, M Mayer, F Liebsch, G Multhaup, S Chiantia (2017)
Direct evidence of amyloid precursor-like protein 1 trans interactions in cell-cell adhesion platforms investigated via fluorescence fluctuation spectroscopyMol Biol Cell, 28
O Krichevsky, G Bonnet (2002)
Fluorescence correlation spectroscopy: the technique and its applicationsReports Prog Phys, 65
J Widengren, P Thyberg (2005)
FCS cell surface measurements – photophysical limitations and consequences on molecular ensembles with heterogenic mobilitiesCytometry A, 68A
SC Weber, AJ Spakowitz, JA Theriot (2012)
Nonthermal ATP-dependent fluctuations contribute to the in vivo motion of chromosomal lociPNAS, 109
N Gröner, J Capoulade, C Cremer, M Wachsmuth (2010)
Measuring and imaging diffusion with multiple scan speed image correlation spectroscopyOpt Express, 18
S Ito, T Sugiyama, N Toitani, G Katayama, H Miyasaka (2007)
Application of fluorescence correlation spectroscopy to the measurement of local temperature in solutions under optical trapping conditionsJ Phys Chem, 111
Fluorescence correlation spectroscopy (FCS) is a powerful tool to quantitatively study the diffusion of fluorescently labeled molecules. It allows in principle important questions of macromolecular transport and supramolecular aggregation in living cells to be addressed. However, the crowded environment inside the cells slows diffusion and limits the reservoir of labeled molecules, causing artifacts that arise especially from photobleaching and limit the utility of FCS in these applications. We present a method to compute the time correlation function from weighted photon arrival times, which compensates computationally during the data analysis for the effect of photobleaching. We demonstrate the performance of this method using numerical simulations and experimental data from model solutions. Using this technique, we obtain correlation functions in which the effect of photobleaching has been removed and in turn recover quantitatively accurate mean-square displacements of the fluorophores, especially when deviations from an ideal Gaussian excitation volume are accounted for by using a reference calibration correlation function. This allows quantitative FCS studies of transport processes in challenging environments with substantial photobleaching like in living cells in the future.
Journal of Fluorescence – Springer Journals
Published: Jan 24, 2018
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