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

Rice University, Houston, Texas, United States
Journal of Biomedical Optics (Impact Factor: 2.86). 11/2006; 11(6):064013. DOI: 10.1117/1.2372457
Source: PubMed


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 100 nm 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 100 nm.

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    • "A standing wave embodiment of this technology has been Correspondence to: Pascal Vallotton. Tel: +61 (0)2 9325 3208; fax: +61 (0)2 9325 3200; e-mail: reported that allows improving the lateral resolution down to 100 nm (Gliko et al., 2006). Gordon et al. measure nanometre range distances between two single identical fluorophores by relying on the analysis of the Airy-like intensity patterns, before and subsequent to the photo-bleaching of one of the fluorophores (Gordon et al., 2004). "
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