Internet-based image analysis quantifies contractile behavior of individual fibroblasts inside model tissue.
In a cell-populated collagen gel, intrinsic fiber structure visible in differential interference contrast images can provide markers for an in situ strain gauge to quantify cell-gel mechanics, while optical sections of fluorescent protein distribution capture cytoskeletal kinematics. Mechanics quantification can be derived automatically from timelapse differential interference contrast images using a Deformation Quantification and Analysis software package accessible online at http://dqa.web.cmu.edu. In our studies, fibroblast contractile machinery was observed to function entirely within pseudopods, while GFP-alpha-actinin concentrated in pseudopod tips and cortex. Complex strain patterns around individual cells showed instances of both elastic and inelastic strain transmission, suggesting a role in observed long-range alignment of cells.
Vanni
Steven
S
Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, USA.
Lagerholm
B Christoffer
BC
Otey
Carol
C
Taylor
D Lansing
DL
Lanni
Frederick
F
D016475:3T3 Cells / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D000818:Animals; D032701:Biomimetics / Q000379:methods; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D003094:Collagen / Q000502:physiology* / Q000648:ultrastructure*; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D015638:Cytochalasin D / Q000494:pharmacology; D004548:Elasticity; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D020407:Internet; D040542:Mechanotransduction, Cellular / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009038:Motion; D015739:Nocodazole / Q000494:pharmacology; D012984:Software; D013314:Stress, Mechanical
C093679:Vitrogen; D015638:Cytochalasin D; D003094:Collagen; D015739:Nocodazole