Automated Analysis of Cell-Matrix Adhesions in 2D and 3D Environments

• Published in: Scientific reports Vol. 5; no. 1; p. 8124
• Main Authors: Broussard, Joshua A., Diggins, Nicole L., Hummel, Stephen, Georgescu, Walter, Quaranta, Vito, Webb, Donna J.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 29-01-2015; Nature Publishing Group
• Subjects: 14; 14/19; 14/35; 631/1647/328; 631/80/79/2027; Adhesion; Algorithms; Animals; Automation; Cell adhesion & migration; Cell Culture Techniques - methods; Cell Line, Tumor; Cell migration; Cell survival; Cell-Matrix Junctions - drug effects; Collagen Type I - pharmacology; Green Fluorescent Proteins - metabolism; Humanities and Social Sciences; Humans; Image Processing, Computer-Assisted; Molecular modelling; Morphogenesis; multidisciplinary; Paxillin - metabolism; Rats; Reproducibility of Results; Science; Segmentation; Time-Lapse Imaging; Tumorigenesis; Wound healing
• ISSN: 2045-2322; 2045-2322
• 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
p
latform for the
a
utomated
a
nalysis,
s
egmentation and
t
racking of
a
dhesions (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.