Quantifying Molecular Dynamics within Complex Cellular Morphologies using LLSM-FRAP.
FRAP
actin cytoskeleton
diffusion
lattice light sheet microscopy
membranes
Journal
Small methods
ISSN: 2366-9608
Titre abrégé: Small Methods
Pays: Germany
ID NLM: 101724536
Informations de publication
Date de publication:
06 2022
06 2022
Historique:
received:
01
02
2022
pubmed:
29
3
2022
medline:
18
6
2022
entrez:
28
3
2022
Statut:
ppublish
Résumé
Quantifying molecular dynamics within the context of complex cellular morphologies is essential toward understanding the inner workings and function of cells. Fluorescence recovery after photobleaching (FRAP) is one of the most broadly applied techniques to measure the reaction diffusion dynamics of molecules in living cells. FRAP measurements typically restrict themselves to single-plane image acquisition within a subcellular-sized region of interest due to the limited temporal resolution and undesirable photobleaching induced by 3D fluorescence confocal or widefield microscopy. Here, an experimental and computational pipeline combining lattice light sheet microscopy, FRAP, and numerical simulations, offering rapid and minimally invasive quantification of molecular dynamics with respect to 3D cell morphology is presented. Having the opportunity to accurately measure and interpret the dynamics of molecules in 3D with respect to cell morphology has the potential to reveal unprecedented insights into the function of living cells.
Identifiants
pubmed: 35344286
doi: 10.1002/smtd.202200149
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2200149Subventions
Organisme : Rosalind Franklin Institute and the Kennedy Trust for Rheumatology Research
ID : 212343/Z/18/Z
Organisme : Rosalind Franklin Institute and the Kennedy Trust for Rheumatology Research
ID : EP/S004459/1
Informations de copyright
© 2022 The Authors. Small Methods published by Wiley-VCH GmbH.
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