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Research Papers

Atomic force-multi-optical imaging integrated microscope for monitoring molecular dynamics in live cells

[+] Author Affiliations
Andreea Trache, Gerald A. Meininger

Texas A&M University System Health Science Center, Department of Medical Physiology, Cardiovascular Research Institute, College of Medicine, College Station, Texas 77843-1114

J. Biomed. Opt. 10(6), 064023 (December 29, 2005). doi:10.1117/1.2146963
History: Received March 23, 2005; Revised June 21, 2005; Accepted June 22, 2005; Published December 29, 2005
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A novel hybrid imaging system is constructed integrating atomic force microscopy (AFM) with a combination of optical imaging techniques that offer high spatial resolution. The main application of this instrument (the NanoFluor microscope) is the study of mechanotransduction with an emphasis on extracellular matrix-integrin-cytoskeletal interactions and their role in the cellular responses to changes in external chemical and mechanical factors. The AFM allows the quantitative assessment of cytoskeletal changes, binding probability, adhesion forces, and micromechanical properties of the cells, while the optical imaging applications allow thin sectioning of the cell body at the coverslip-cell interface, permitting the study of focal adhesions using total internal reflection fluorescence (TIRF) and internal reflection microscopy (IRM). Combined AFM-optical imaging experiments show that mechanical stimulation at the apical surface of cells induces a force-generating cytoskeletal response, resulting in focal contact reorganization on the basal surface that can be monitored in real time. The NanoFluor system is also equipped with a novel mechanically aligned dual camera acquisition system for synthesized Forster resonance energy transfer (FRET). The integrated NanoFluor microscope system is described, including its characteristics, applications, and limitations.

Figures in this Article
© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Andreea Trache and Gerald A. Meininger
"Atomic force-multi-optical imaging integrated microscope for monitoring molecular dynamics in live cells", J. Biomed. Opt. 10(6), 064023 (December 29, 2005). ; http://dx.doi.org/10.1117/1.2146963


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