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Focusing on TIRF Microscopy

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Bill Fester, Olympus America Inc.

Applications from cell dynamics and single-molecule events to vesicle and protein tracking are all within the realm of total internal reflection fluorescence (TIRF). The technique is so named because it involves total internal reflection of the excitation light. The method requires no X-, Y- or Z-stage motorization because the cells are stationary. The evanescent wave in the illumination extends only about 100 nm from the coverslip/ water interface, so any area of interest must be within that region. That means growing cells on a coverslip. If one simply places a coverslip on a cell to...Read full article

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    Published: July 2003
    Glossary
    total internal reflection fluorescence
    Total internal reflection fluorescence (TIRF) microscopy is an advanced fluorescence microscopy technique that leverages the principle of total internal reflection to selectively illuminate and image a thin section of a specimen near the interface of two media, typically a glass coverslip and the sample medium. TIRF microscopy allows for high-resolution imaging of fluorescently labeled structures near the cell membrane or other interfaces, offering improved signal-to-noise ratios and reduced...
    Basic Sciencecell dynamicsFeaturesMicroscopyprotein trackingsingle-molecule eventsTIRFtotal internal reflection fluorescencevesicle

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