Research Papers

Integrated microscopy for real-time imaging of mechanotransduction studies in live cells

[+] Author Affiliations
Andreea Trache

Texas A&M Health Science Center, Cardiovascular Research Institute, College of Medicine, Department of Systems Biology and Translational Medicine, College Station, Texas 77843-1114 and Texas A&M University, Department of Biomedical Engineering, College Station, Texas 77840

Soon-Mi Lim

Texas A&M Health Science Center, Cardiovascular Research Institute, College of Medicine, Department of Systems Biology and Translational Medicine, College Station, Texas 77843-1114

J. Biomed. Opt. 14(3), 034024 (June 30, 2009). doi:10.1117/1.3155517
History: Received October 02, 2008; Revised April 02, 2009; Accepted April 04, 2009; Published June 30, 2009
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Mechanical force is an important stimulus and determinant of many vascular smooth muscle cell functions including contraction, proliferation, migration, and cell attachment. Transmission of force from outside the cell through focal adhesions controls the dynamics of these adhesion sites and initiates intracellular signaling cascades that alter cellular behavior. To understand the mechanism by which living cells sense mechanical forces, and how they respond and adapt to their environment, a critical first step is to develop a new technology to investigate cellular behavior at subcellular level that integrates an atomic force microscope (AFM) with total internal reflection fluorescence (TIRF) and fast-spinning disk (FSD) confocal microscopy, providing high spatial and temporal resolution. AFM uses a nanosensor to measure the cell surface topography and can apply and measure mechanical force with high precision. TIRF microscopy is an optical imaging technique that provides high-contrast images with high z-resolution of fluorescently labeled molecules in the immediate vicinity of the cell–coverslip interface. FSD confocal microscopy allows rapid 3-D imaging throughout the cell in real time. The integrated system is broadly applicable across a wide range of molecular dynamic studies in any adherent live cells, allowing direct optical imaging of cell responses to mechanical stimulation in real time.

Figures in this Article
© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Andreea Trache and Soon-Mi Lim
"Integrated microscopy for real-time imaging of mechanotransduction studies in live cells", J. Biomed. Opt. 14(3), 034024 (June 30, 2009). ; http://dx.doi.org/10.1117/1.3155517


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