Advanced quantification for single-cell adhesion by variable-angle TIRF nanoscopy.

Over the last decades, several techniques have been developed to study cell adhesion; however, they present significant shortcomings. Such techniques mostly focus on strong adhesion related to specific protein-protein associations, such as ligand-receptor binding in focal adhesions. Therefore, weak adhesion, related to less specific or nonspecific cell-substrate interactions, are rarely addressed.

Incidence angle calibration for prismless total internal reflection fluorescence microscopy.

We propose a calibration routine useful to evaluate the incident angle in total internal reflection fluorescence (TIRF) microscopy. This procedure is based on critical angle measurements conducted in the back focal plane (BFP) of the objective. Such BFP imaging can be easily implemented on any TIRF setup, making this technique very attractive.