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An Alternative Framework for Fluorescence Correlation Spectroscopy

Authors :
Sanjeevi Sivasankar
Ioannis Sgouralis
Omer Shafraz
Steve Pressé
Sina Jazani
Source :
Biophysical Journal. 116:282a
Publication Year :
2019
Publisher :
Elsevier BV, 2019.

Abstract

Fluorescence correlation spectroscopy (FCS), is a flexible and widely used tool routinely exploited in vivo. While FCS provides estimates of dynamical quantities, such as diffusion coefficients, it demands high molecular concentrations, high signal to noise ratio and long time traces typically in the minute range. This is by contrast to data acquisition which, in principle, provides information on the micro second timescales that has, thus far, evaded analysis. To overcome these limitations, we adapt novel tools inspired by Bayesian non-parametrics. Using this framework, which starts from the direct analysis of photon counts, we achieve the same accuracy as FCS, but with much shorter traces, even at nearly single molecule concentrations. Our new analysis extends the capability of confocal microscopy based approaches, to probe processes several orders of magnitude faster in time and permits a reduction of phototoxic effects on living samples induced by long periods of light exposure.

Details

ISSN :
00063495
Volume :
116
Database :
OpenAIRE
Journal :
Biophysical Journal
Accession number :
edsair.doi...........771e2bed5715cf3a7a1facb69e0138ff