Large-field-of-view imaging by multi-pupil adaptive optics

Abstract

For in vivo deep imaging at high spatiotemporal resolutions, we developed Multi-Pupil Adaptive Optics (MPAO) which enables simultaneous wavefront correction over a large imaging field-of-view. The current implementation improves correction area by nine times over that of conventional methods. MPAO’s capability of spatially independent wavefront control further enables 3D nonplanar imaging. We applied MPAO to in vivo structural and functional imaging of biological dynamics in mammalian brain.

High contrast imaging and flexible photomanipulation for quantitative in vivo multiphoton imaging with polygon scanning microscope

“…In conclusion, the new polygon microscope displays the highest SBP at video rate that when combined with photomanipulation capabilities can achieve a greater breadth of quantitative in vivo imaging that was not possible before in polygon scanning mirrors. Furthermore, this method can also open up multi‐FOV adaptive optics corrections or localized super‐resolution microscopy .…”

Perspective: Wavefront shaping techniques for controlling multiple light scattering in biological tissues: Toward in vivo applications

“…127 Instead of dividing the aberrating volume into different layers, the aberrations that differ for different parts of the field of view were measured and corrected for independently in a parallel manner. As previously discussed, image-based wavefront sensing already provides us with the information for correctly applying different corrections to different parts of the field of view.…”

Multi-plane Imaging of Neural Activity From the Mammalian Brain Using a Fast-switching Liquid Crystal Spatial Light Modulator

“…As a result, the microscope can focus at different axial planes by applying different defocus wavefront patterns. Before each imaging session, adaptive optics procedures were performed on 2 µm fluorescent beads to correct for residual optical aberrations due to possible slight misalignments using a procedure described in [22], ensuring we are applying ideal defocus wavefront patterns in the system. Updating the defocus patterns on the SLM was synchronized to a single-frame resonant-galvo scan for image acquisition by a handshake protocol.…”

“…As shown in Fig.4, with an effective NA of 0.65 but without adaptive optics, the PSF degraded quickly when the SLM was used to focus tens of microns away from the nominal focal plane. When adaptive optics procedures as described in [22] were applied on top of different defocus wavefront patterns, the PSFs at different depths were recovered over an axial range of ∼ 100 µm.…”