TY  - JOUR
A1  - Khadem, S. M. J.
A1  - Hille, Carsten
A1  - Löhmannsröben, Hans-Gerd
A1  - Sokolov, Igor M.
T1  - Spot variation fluorescence correlation spectroscopy by data post-processing
T2  - Scientific reports
N2  - Spot variation fluorescence correlation spectroscopy (SV-FCS) is a variant of the FCS techniques which may give useful information about the structural organisation of the medium in which the diffusion takes place. We show that the same results can be obtained by post-processing the photon count data from ordinary FCS measurements. By using this method, one obtains the fluorescence autocorrelation functions for sizes of confocal volume, which are effectively smaller than that of the initial FCS measurement. The photon counts of the initial experiment are first transformed into smooth intensity trace using kernel smoothing method or to a piecewise-continuous intensity trace using binning and then a non-linear transformation is applied to this trace. The result of this transformation mimics the photon count rate in an experiment performed with a smaller confocal volume. The applicability of the method is established in extensive numerical simulations and directly supported in in-vitro experiments. The procedure is then applied to the diffusion of AlexaFluor647-labeled streptavidin in living cells.
Y1  - 2017
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/46524
SN  - 2045-2322
VL  - 7
SP  - 1
EP  - 9
PB  - Nature Publ. Group
CY  - London
ER  -