TY - JOUR
T1 - Automated analysis of cell-matrix adhesions in 2D and 3D environments
AU - Broussard, Joshua A.
AU - Diggins, Nicole L.
AU - Hummel, Stephen
AU - Georgescu, Walter
AU - Quaranta, Vito
AU - Webb, Donna J.
N1 - Funding Information:
We thank Susan Craig and Rick Horwitz for generously providing us with reagents. This work was supported by National Institutes of Health (NIH) grants GM092914 and by National Center for Research Resources Grant S10RR025524 to D.J.W. J.A.B. was supported by predoctoral training grant CA078136 from NIH.
PY - 2015/1
Y1 - 2015/1
N2 - 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.
AB - 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.
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U2 - 10.1038/srep08124
DO - 10.1038/srep08124
M3 - Article
C2 - 25630460
AN - SCOPUS:84923090509
VL - 5
SP - 8124
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
M1 - 8124
ER -