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TOTAL INTERNAL REFLECTION FLUORESCENCE (TIRF)
Imaging of dense-core vesicle release from bovine Chromaffin cell taken on Olympus IX81 system. Release of GFP-labelled vesicles is heralded with intensity increase and followed by diffusion of marker.
Credits; LiMa, Lou Philipson, Aaron Fox, Vytas Bindokas (1-13-04)
TIRF is used to observe the surface regions of cells or the fluorescence of a single molecule without interference from possible background fluorescence. Evanescent waves selectively excite fluorophores from a small region (less than 200 nm). The confocal facility has the Olympus high numerical aperture apochromatic objective matched for total internal refllection fluorescence microscopy. The reflective index of the objective is 1.78. Special immersion oil and coversips match the reflective index.
