Please use this identifier to cite or link to this item: https://hdl.handle.net/1959.11/23305
Title: Quantitative image mean squared displacement (iMSD) analysis of the dynamics of profilin 1 at the membrane of live cells
Contributor(s): Davey, Rhonda (author); Digman, Michelle (author); Gratton, Enrico (author); Moens, Pierre (author)orcid 
Publication Date: 2018
DOI: 10.1016/j.ymeth.2017.12.002
Handle Link: https://hdl.handle.net/1959.11/23305
Abstract: Image mean square displacement analysis (iMSD) is a method allowing the mapping of diffusion dynamics of molecules in living cells. However, it can also be used to obtain quantitative information on the diffusion processes of fluorescently labelled molecules and how their diffusion dynamics change when the cell environment is modified. In this paper, we describe the use of iMSD to obtain quantitative data of the diffusion dynamics of a small cytoskeletal protein, profilin 1 (pfn1), at the membrane of live cells and how its diffusion is perturbed when the cells are treated with Cytochalasin D and/or the interactions of pfn1 are modified when its actin and polyphosphoinositide binding sites are mutated (pfn1-R88A). Using total internal reflection fluorescence microscopy images, we obtained data on isotropic and confined diffusion coefficients, the proportion of cell areas where isotropic diffusion is the major diffusion mode compared to the confined diffusion mode, the size of the confinement zones and the size of the domains of dynamic partitioning of pfn1. Using these quantitative data, we could demonstrate a decreased isotropic diffusion coefficient for the cells treated with Cytochalasin D and for the pfn1-R88A mutant. We could also see changes in the modes of diffusion between the different conditions and changes in the size of the zones of pfn1 confinements for the pfn1 treated with Cytochalasin D. All of this information was acquired in only a few minutes of imaging per cell and without the need to record thousands of single molecule trajectories.
Publication Type: Journal Article
Source of Publication: Methods, v.140-141, p. 119-125
Publisher: Academic Press
Place of Publication: United States of America
ISSN: 1095-9130
1046-2023
Field of Research (FOR): 111201 Cancer Cell Biology
060103 Cell Development, Proliferation and Death
029901 Biological Physics
Peer Reviewed: Yes
HERDC Category Description: C1 Refereed Article in a Scholarly Journal
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Appears in Collections:Journal Article
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