Automated analysis of cell-matrix adhesions in 2D and 3D environments.

Broussard JA, Diggins NL, Hummel S, Georgescu W, Quaranta V, Webb DJ
Sci Rep. 2015 5: 8124

PMID: 25630460 · PMCID: PMC4309964 · DOI:10.1038/srep08124

Cell-matrix adhesions are of great interest because of their contribution to numerous biological processes, including cell migration, differentiation, proliferation, survival, tissue morphogenesis, wound healing, and tumorigenesis. Adhesions are dynamic structures that are classically defined on two-dimensional (2D) substrates, though the need to analyze adhesions in more physiologic three-dimensional (3D) environments is being increasingly recognized. However, progress has been greatly hampered by the lack of available tools to analyze adhesions in 3D environments. To address this need, we have developed a platform for the automated analysis, segmentation, and tracking of adhesions (PAASTA) based on an open source MATLAB framework, CellAnimation. PAASTA enables the rapid analysis of adhesion dynamics and many other adhesion characteristics, such as lifetime, size, and location, in 3D environments and on traditional 2D substrates. We manually validate PAASTA and utilize it to quantify rate constants for adhesion assembly and disassembly as well as adhesion lifetime and size in 3D matrices. PAASTA will be a valuable tool for characterizing adhesions and for deciphering the molecular mechanisms that regulate adhesion dynamics in 3D environments.

MeSH Terms (14)

Algorithms Animals Automation Cell-Matrix Junctions Cell Culture Techniques Cell Line, Tumor Collagen Type I Green Fluorescent Proteins Humans Image Processing, Computer-Assisted Paxillin Rats Reproducibility of Results Time-Lapse Imaging

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