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The extracellular matrix or ECM holds cells together to form a tissue and allows the cells within the tissue to communicate. ECM comprises proteins such as fibronectin, collagen, laminin, etc. The most abundant protein in this space is collagen. Collagen fibers are interwoven with carbohydrate-containing protein molecules called proteoglycans. ECM allows cell migration and provides a structural scaffold at cell adhesion that anchors the cell when the extracellular matrix proteins interact with...

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Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts

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Published on: August 27, 2012

Quantitative multicolor compositional imaging resolves molecular domains in cell-matrix adhesions.

Eli Zamir¹, Benjamin Geiger, Zvi Kam

¹Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot, Israel.

|April 3, 2008

Summary

Compositional imaging reveals molecular signatures within cell adhesion sites. This advanced microscopy technique maps complex molecular assemblies and their variations, offering new insights into cellular structures.

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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

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Published on: December 9, 2013

Area of Science:

Cellular biology
Molecular imaging
Biophysics

Background:

Cellular processes occur in dynamic, multi-molecular compartments requiring high spatio-temporal resolution.
Cell-matrix adhesion sites, composed of cytoskeletal and signaling proteins, are crucial but poorly defined molecularly.
Variability in morphology, dynamics, and function of adhesions highlights the need for detailed molecular characterization.

Purpose of the Study:

To develop and apply a compositional imaging approach for analyzing multi-component molecular compositions in cells.
To resolve molecular complexes within focal adhesions and their response to Rho-kinase inhibition.
To enhance spatial information with content-resolved dimensions for studying cellular assemblies.

Main Methods:

Utilized microscopy-acquired multicolor data.
Applied multi-dimensional clustering of pixels based on composition similarity.
Displayed cellular distribution of identified composition clusters.

Main Results:

Resolved molecular complexes associated with focal adhesions.
Identified compositional variations between and along individual adhesion sites.
Revealed distinct sensitivities of focal adhesion complexes to Rho-kinase inhibition.

Conclusions:

Multicolor compositional imaging defines molecular signatures of focal adhesions and their sub-domains.
This approach adds content-resolved dimensions to spatial information.
Compositional imaging is a powerful tool for studying and mapping multi-molecular assemblies at sub-micron resolution.