Imaging performance of microscopy adaptive-optics system using scene-based wavefront sensing.

Abstract

.Significance: A scene-based adaptive-optics (AO) system is developed and a method for investigating its imaging performance is proposed. The system enables derivation of Strehl ratios from observed images via collaboration with computer simulations. The resultant Strehl ratios are comparable with those of other current AO systems.Aim: For versatile and noninvasive AO microscopy, a scene-based wavefront-sensing technique working on a Shack–Hartmann wavefront sensor is developed in a modal control system. The purpose of the research is to clarify the imaging performance of the AO system via the derivation of Strehl ratios from observed images toward applications in microscopy of living cells and tissues.Approach: Two imaging metrics that can be directly measured from observed images (i.e., an energy concentration ratio and unbiased maximum ratio) are defined and related to the Strehl ratio via computer simulations. Experiments are conducted using artificial targets to measure the imaging metrics, which are then converted to Strehl ratios.Results: The resultant Strehl ratios are and 0.5 in the cases of defocus and higher aberrations, respectively. The half-widths at half-maximum of the AO-corrected bead images are favorably comparable to those of on-focus images under simple defocus aberration, and the AO system works both under bright-field illumination and on fluorescent bead images.Conclusions: The proposed scene-based AO system is expected to work with a Strehl ratio of more than 0.5 when applied to high-resolution live imaging of cells and tissues under bright-field and fluorescence microscopies.