Total Internal Reflection Fluorescence (TIRF) Microscopy

Abstract

TIRF microscopy is a technique for imaging the surface of cells that utilizes a thin field of illumination to image fluorophores on or near to the coverslip. TIRF is a highly sensitive widefield camera-based technique providing an excellent signal-to-noise ratio compared with epi-fluorescence or confocal imaging methods. TIRF can be used alone or in combination with other imaging techniques to give an “inside and out” view of a specimen. This technical note describes the theoretical basis of TIRF microscopy, the hardware required and examples of TIRF applications.