TIRF Microscopy highlights events at and near the plasma membrane

Edited by March 8, 2019


Communicated by the Biology and Pathology of the Endocrine Pancreas Team

The TIRF (Total Internal Reflection Fluorescence) technique uses the unique properties of an evanescent wave to excitate and selectively illuminate fluorophores located in a restricted area of ​​the sample, adjacent to the glass–cell interface. The principle of obtaining an evanescent wave is as follows: when a laser beam is directed towards the glass slide at an incident angle greater than a critical angle, the beam is not then refracted but totally reflected and induces the formation of a light wave, called evanescent wave, in the cell adjacent to the glass-cell interface. The penetration depth of this evanescent wave typically ranges can go up to 200 nm. The plasma membrane of an adherent cell lies well within the region of excitation, allowing imaging of processes occurring at or near the membrane.


Disrupted dynamics of F-actin and insulin granule fusion in INS-1 832/13 beta-cells exposed to glucotoxicity: partial restoration by glucagon-like peptide 1. Quinault A, Gausseres B, Baibe D, Chebbah N, Portha B, Movassat J, Tourrel-Cuzin C. Biochim. Biophys. Acta. (2016), 182:1401-11.


TIRF facility