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Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions

BACKGROUND: Cellular processes occur within dynamic and multi-molecular compartments whose characterization requires analysis at high spatio-temporal resolution. Notable examples for such complexes are cell-matrix adhesion sites, consisting of numerous cytoskeletal and signaling proteins. These adhe...

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Detalles Bibliográficos
Autores principales: Zamir, Eli, Geiger, Benjamin, Kam, Zvi
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2270910/
https://www.ncbi.nlm.nih.gov/pubmed/18382676
http://dx.doi.org/10.1371/journal.pone.0001901
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author Zamir, Eli
Geiger, Benjamin
Kam, Zvi
author_facet Zamir, Eli
Geiger, Benjamin
Kam, Zvi
author_sort Zamir, Eli
collection PubMed
description BACKGROUND: Cellular processes occur within dynamic and multi-molecular compartments whose characterization requires analysis at high spatio-temporal resolution. Notable examples for such complexes are cell-matrix adhesion sites, consisting of numerous cytoskeletal and signaling proteins. These adhesions are highly variable in their morphology, dynamics, and apparent function, yet their molecular diversity is poorly defined. METHODOLOGY/PRINCIPAL FINDINGS: We present here a compositional imaging approach for the analysis and display of multi-component compositions. This methodology is based on microscopy-acquired multicolor data, multi-dimensional clustering of pixels according to their composition similarity and display of the cellular distribution of these composition clusters. We apply this approach for resolving the molecular complexes associated with focal-adhesions, and the time-dependent effects of Rho-kinase inhibition. We show here compositional variations between adhesion sites, as well as ordered variations along the axis of individual focal-adhesions. The multicolor clustering approach also reveals distinct sensitivities of different focal-adhesion-associated complexes to Rho-kinase inhibition. CONCLUSIONS/SIGNIFICANCE: Multicolor compositional imaging resolves “molecular signatures” characteristic to focal-adhesions and related structures, as well as sub-domains within these adhesion sites. This analysis enhances the spatial information with additional “contents-resolved” dimensions. We propose that compositional imaging can serve as a powerful tool for studying complex multi-molecular assemblies in cells and for mapping their distribution at sub-micron resolution.
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spelling pubmed-22709102008-04-02 Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions Zamir, Eli Geiger, Benjamin Kam, Zvi PLoS One Research Article BACKGROUND: Cellular processes occur within dynamic and multi-molecular compartments whose characterization requires analysis at high spatio-temporal resolution. Notable examples for such complexes are cell-matrix adhesion sites, consisting of numerous cytoskeletal and signaling proteins. These adhesions are highly variable in their morphology, dynamics, and apparent function, yet their molecular diversity is poorly defined. METHODOLOGY/PRINCIPAL FINDINGS: We present here a compositional imaging approach for the analysis and display of multi-component compositions. This methodology is based on microscopy-acquired multicolor data, multi-dimensional clustering of pixels according to their composition similarity and display of the cellular distribution of these composition clusters. We apply this approach for resolving the molecular complexes associated with focal-adhesions, and the time-dependent effects of Rho-kinase inhibition. We show here compositional variations between adhesion sites, as well as ordered variations along the axis of individual focal-adhesions. The multicolor clustering approach also reveals distinct sensitivities of different focal-adhesion-associated complexes to Rho-kinase inhibition. CONCLUSIONS/SIGNIFICANCE: Multicolor compositional imaging resolves “molecular signatures” characteristic to focal-adhesions and related structures, as well as sub-domains within these adhesion sites. This analysis enhances the spatial information with additional “contents-resolved” dimensions. We propose that compositional imaging can serve as a powerful tool for studying complex multi-molecular assemblies in cells and for mapping their distribution at sub-micron resolution. Public Library of Science 2008-04-02 /pmc/articles/PMC2270910/ /pubmed/18382676 http://dx.doi.org/10.1371/journal.pone.0001901 Text en Zamir et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zamir, Eli
Geiger, Benjamin
Kam, Zvi
Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title_full Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title_fullStr Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title_full_unstemmed Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title_short Quantitative Multicolor Compositional Imaging Resolves Molecular Domains in Cell-Matrix Adhesions
title_sort quantitative multicolor compositional imaging resolves molecular domains in cell-matrix adhesions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2270910/
https://www.ncbi.nlm.nih.gov/pubmed/18382676
http://dx.doi.org/10.1371/journal.pone.0001901
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