[Basics of TIRFM].
TIRFM (total internal reflection fluorescence microscopy) is a fast-growing imaging technique which is currently getting significant attention of scientific community. The great advantage of this type of microscopy is in its optical sectioning capability superior to other techniques, such as confocal microscopy. Better signal to noise ratio makes it possible to directly observe the behavior of biomolecules at the single-molecule level both in vitro and in vivo. In this review, I will elucidate the physical theory of the TIR and optical sectioning effects and will provide with the examples of how they can be utilized in microscopy and bioscience. Finally, I will go through an example of a design procedure of a home-built objective-type TIRFM and demonstrate the latest single molecule imaging example acquired by our TIRFM setup.
Nishikawa
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JST CREST Soft Nano Machine.
D008856:Microscopy, Fluorescence / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*