Research Papers

Standing wave total internal reflection fluorescence microscopy to measure the size of nanostructures in living cells

[+] Author Affiliations
Olga Gliko

Baylor College of Medicine, Department of Otolaryngology—Head & Neck Surgery, One Baylor Plaza, NA516, Houston, Texas 77030

Gaddum D. Reddy, Bahman Anvari

Rice University, Department of Bioengineering, Houston, Texas 77005

William E. Brownell

Baylor College of Medicine, Department of Otolaryngology—Head & Neck Surgery, One Baylor Plaza, NA516, Houston, Texas 77030

Peter Saggau

Baylor College of Medicine, Department of Neuroscience, One Baylor Plaza, S603, Houston, Texas 77030

J. Biomed. Opt. 11(6), 064013 (November 14, 2006). doi:10.1117/1.2372457
History: Received April 10, 2006; Revised August 29, 2006; Accepted August 31, 2006; Published November 14, 2006; Online November 14, 2006
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We present the first application of standing wave fluorescence microscopy (SWFM) to determine the size of biological nanostructures in living cells. The improved lateral resolution of less than 100nm enables superior quantification of the size of subcellular structures. We demonstrate the ability of SWFM by measuring the diameter of biological nanotubes (membrane tethers formed between cells). The combination of SWFM with total internal reflection (TIR), referred to as SW-TIRFM, allows additional improvement of axial resolution by selective excitation of fluorescence in a layer of about 100nm.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

Olga Gliko ; Gaddum D. Reddy ; Bahman Anvari ; William E. Brownell and Peter Saggau
"Standing wave total internal reflection fluorescence microscopy to measure the size of nanostructures in living cells", J. Biomed. Opt. 11(6), 064013 (November 14, 2006). ; http://dx.doi.org/10.1117/1.2372457


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