Spot variation fluorescence correlation spectroscopy by data post-processing

Abstract 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...

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Autores principales:	S. M. J. Khadem, C. Hille, H.-G. Löhmannsröben, I. M. Sokolov
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3d3556e508a74bbbbfdf29276e8bb738
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spelling	oai:doaj.org-article:3d3556e508a74bbbbfdf29276e8bb7382021-12-02T11:52:24ZSpot variation fluorescence correlation spectroscopy by data post-processing10.1038/s41598-017-05672-82045-2322https://doaj.org/article/3d3556e508a74bbbbfdf29276e8bb7382017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05672-8https://doaj.org/toc/2045-2322Abstract 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.S. M. J. KhademC. HilleH.-G. LöhmannsröbenI. M. SokolovNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q S. M. J. Khadem C. Hille H.-G. Löhmannsröben I. M. Sokolov Spot variation fluorescence correlation spectroscopy by data post-processing
description	Abstract 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.
format	article
author	S. M. J. Khadem C. Hille H.-G. Löhmannsröben I. M. Sokolov
author_facet	S. M. J. Khadem C. Hille H.-G. Löhmannsröben I. M. Sokolov
author_sort	S. M. J. Khadem
title	Spot variation fluorescence correlation spectroscopy by data post-processing
title_short	Spot variation fluorescence correlation spectroscopy by data post-processing
title_full	Spot variation fluorescence correlation spectroscopy by data post-processing
title_fullStr	Spot variation fluorescence correlation spectroscopy by data post-processing
title_full_unstemmed	Spot variation fluorescence correlation spectroscopy by data post-processing
title_sort	spot variation fluorescence correlation spectroscopy by data post-processing
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/3d3556e508a74bbbbfdf29276e8bb738
work_keys_str_mv	AT smjkhadem spotvariationfluorescencecorrelationspectroscopybydatapostprocessing AT chille spotvariationfluorescencecorrelationspectroscopybydatapostprocessing AT hglohmannsroben spotvariationfluorescencecorrelationspectroscopybydatapostprocessing AT imsokolov spotvariationfluorescencecorrelationspectroscopybydatapostprocessing
_version_	1718395041545715712

Spot variation fluorescence correlation spectroscopy by data post-processing

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