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  5. Advanced processing and analysis of conventional confocal microscopy generated scanning FCS data
 
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Advanced processing and analysis of conventional confocal microscopy generated scanning FCS data
File(s)
163766v2.full.pdf (1.49 MB)
Working paper
Author(s)
Waithe, Dominic
Schneider, Falk
Chojnacki, Jakub
Clausen, Mathias
Shrestha, Dilip
more
Type
Working Paper
Abstract
Scanning Fluorescence Correlation Spectroscopy (scanning FCS) is a variant of conventional point FCS that allows molecular diffusion at multiple locations to be measured simultaneously. It enables disclosure of potential spatial heterogeneity in molecular diffusion dynamics and also the acquisition of a large amount of FCS data at the same time, providing large statistical accuracy. Here, we optimize the processing and analysis of these large-scale acquired sets of FCS data. On one hand we present FoCuS-scan, scanning FCS software that provides an end-to-end solution for processing and analysing scanning data acquired on commercial turnkey confocal systems. On the other hand, we provide a thorough characterisation of large-scale scanning FCS data over its intended time-scales and applications and propose a unique solution for the bias and variance observed when studying slowly diffusing species. Our manuscript enables researchers to straightforwardly utilise scanning FCS as a powerful technique for measuring diffusion across a broad range of physiologically relevant length scales without specialised hardware or expensive software.
Date Issued
2017-07-31
Citation
2017
URI
http://hdl.handle.net/10044/1/85235
DOI
https://www.dx.doi.org/10.1101/163766
ISSN
1046-2023
Publisher
bioRxiv
Copyright Statement
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
License URL
https://creativecommons.org/licenses/by-nc-nd/4.0/
Sponsor
Marie Curie Career Integration Grant
Grant Number
PCIG13-GA-2013-618914
Subjects
1103 Clinical Sciences
Publication Status
Published
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