Raster Image Correlation Spectroscopy Performance Evaluation.
Journal
Biophysical journal
ISSN: 1542-0086
Titre abrégé: Biophys J
Pays: United States
ID NLM: 0370626
Informations de publication
Date de publication:
19 11 2019
19 11 2019
Historique:
received:
17
04
2019
revised:
28
08
2019
accepted:
30
09
2019
pubmed:
2
11
2019
medline:
24
9
2020
entrez:
1
11
2019
Statut:
ppublish
Résumé
Raster image correlation spectroscopy (RICS) is a fluorescence image analysis method for extracting the mobility, concentration, and stoichiometry of diffusing fluorescent molecules from confocal image stacks. The method works by calculating a spatial correlation function for each image and analyzing the average of those by model fitting. Rules of thumb exist for RICS image acquisitioning, yet a rigorous theoretical approach to predict the accuracy and precision of the recovered parameters has been lacking. We outline explicit expressions to reveal the dependence of RICS results on experimental parameters. In terms of imaging settings, we observed that a twofold decrease of the pixel size, e.g., from 100 to 50 nm, decreases the error on the translational diffusion constant (D) between three- and fivefold. For D = 1 μm
Identifiants
pubmed: 31668746
pii: S0006-3495(19)30852-5
doi: 10.1016/j.bpj.2019.09.045
pmc: PMC7018992
pii:
doi:
Substances chimiques
enhanced green fluorescent protein
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1900-1914Informations de copyright
Copyright © 2019 Biophysical Society. Published by Elsevier Inc. All rights reserved.
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