A framework for multi-angle TIRF microscope calibration

This communication presents a pipeline for Multi-Angle TIRF calibration from the measurement of the incident angle to the model validation. This problem is of major importance when dealing with 3D reconstruction methods from a set of MA-TIRF acquisitions since the reconstruction accuracy highly depends on the agreement between the theoretical model and the physical system. One main issue is then to build phantom samples with known geometry, or known properties, in order to adjust and/or validate the model. This paper describes such a calibration procedure using a lens as phantom sample and proposes a new model validation experiment based on a dual-color co-localization.