Dynamic imaging of cellular interactions with extracellular matrix
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Adhesive and proteolytic interactions of cells with components of the extracellular matrix (ECM) are fundamental to morphogenesis, tissue assembly and remodeling, and cell migration as well as signal acquisition from tissue-bound factors. The visualization from fixed samples provides snapshot-like, static information on the cellular and molecular dynamics of adhesion receptor and protease functions toward ECM, such as interstitial fibrillar tissues and basement membranes. Recent technological developments additionally support the dynamic imaging of ECM scaffolds and the interaction behavior of cells contained therein. These include differential interference contrast, confocal reflection microscopy, optical coherence tomography, and multiphoton microscopy and second-harmonic generation imaging. Most of these approaches are combined with fluorescence imaging using derivates of GFP and/or other fluorescent dyes. Dynamic 3D imaging has revealed an unexpected degree of dynamics and turnover of cell adhesion and migration as well as basic mechanisms that lead to proteolytic remodeling of connective tissue by stromal cells and invading tumor cells.
KeywordsCollagen matrix Intravital microscopy Cell migration Integrins Attachment Detachment Tissue remodeling Tumor invasion
Movie 1 Time-resolved confocal reflection and fluorescence imaging of MV3 melanoma cell migrating within 3D collagen lattice. Traction of collagen fibers by the cell and forward movement are accompanied by the release of CD44 from the trailing edge upon cell detachment. Cells were labeled with anti-CD44 mAb Hermes-3 and secondary non-cross-linking LRSC-conjugated goat-anti mouse F(ab)’- fragments, thus avoiding receptor hypercrosslinking. Images of the fluorescent (red) and reflection channel (grayscale) were obtained every 5 min and displayed in false-color. Time is indicated in the lower right corner.
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