Modulating phase for adaptive optics and PSF shaping in bio-imaging: Requirements & development of a new deformable mirror tailored to microscopy

Abstract

Modern bio-imaging techniques such as light-sheet, multiphoton and PALM/STORM are now aiming to image more complex biological samples at larger depth and therefore face larger-amplitude and more complex aberrations. We provide an analysis of key requirements driving optimal implementation of adaptive optics (AO) in microscopy, with a focus on wavefront modulators. We show that some specifications of wavefront modulators such as linearity, hysteresis or actuators performance & layout can end up to better AO performance in microscopy systems, when specifically optimized for such use. We then provide design details and characterization results of a newly developed deformable mirror, and report on experimental images obtained from AO-enhanced microscopes based on the device, for several modalities such as light-sheet, multiphoton or super-resolution single molecule localization systems. Finally, we provide recommendations on how to define the right set of AO components, algorithms and overall method depending on modality, instrument and sample constraints.