Evanescent-wave field imaging: an introduction to total internal reflection fluorescence microscopy.

Advancements in technology and computational power in recent years have directly impacted modern microscopy through improvements in light detection, imaging software platforms, as well as integration of complex hardware systems. These successes have allowed for mainstream utilization of previously complex microscopic techniques such as total internal reflection fluorescence (TIRF) microscopy, revealing many aspects of cell biology not previously appreciated. Through the restriction of illumination to areas of cell-coverslip interfaces in combination with modern detectors, TIRF microscopy allows researchers in the life sciences a glimpse of dynamic cellular phenomena with resolutions never before achieved.This chapter provides a basic overview to the concept of TIRF microscopy and some considerations to setting up this technique in the lab.