Wavefront sensing and conjugate adaptive optics in wide-field microscopy
The quality of microscopy imaging is often degraded by sample-induced aberrations. Adaptive optics (AO) is a standard approach to counter such aberrations. In common practice of AO, an active optical correction element, usually a deformable mirror (DM), is usually inserted in the pupil plane of the objective lens, namely pupil AO. However, as first proposed in the astronomy community and now gradually recognized by the optical microscopy community, the placement of the DM in a plane conjugate to a primary sample-induced aberration plane can be more advantageous, especially in situations where the aberration is spatially varying and arises mainly from a dominant layer. We refer to this technique as conjugate AO. In this thesis, we describe two novel implementations of sensor-based conjugate AO in wide-field microscopy, as well as the wavefront sensing techniques we developed for these implementations. Our first implementation is in trans-illumination configuration. The wavefront sensor is based on a technique called partitioned aperture wavefront (PAW) sensing, previously developed in our lab for quantitative phase contrast imaging. Our second conjugate AO is implemented with fluorescence microscopy. The wavefront sensing strategy is based on oblique back-illumination. In both implementations, we addressed the key challenges of developing wavefront sensors that are capable of operating with uncollimated light, which exhibits large diverging angles and may arbitrarily distribute as well. We show that both conjugate AO systems and their wavefront sensors are not only robust, well-suited for video-rate imaging, but also provide large corrected field of view, which is only limited by the microscope itself.