Adaptive Optics Microscopy with Wavefront Sensing Based on Neighbor Correlation.

Complex structures in living cells and tissues induce wavefront errors when light waves pass through them, and images observed with optical microscopes are undesirably blurred. This problem is especially serious for living plant cells because images are strikingly degraded even within a single cell. Adaptive optics (AO) is expected to be a solution to this problem by correcting such wavefront errors, thus enabling high-resolution imaging. In particular, scene-based AO involves wavefront sensing based on the image correlation between subapertures in a Shack-Hartmann wavefront sensor and thus does not require an intense point light source. However, the complex 3D structures of living cells often cause low correlation between subimages, leading to loss of accuracy in wavefront sensing. This paper proposes a novel method for scene-based sensing using only image correlations between adjacent subapertures. The method can minimize changes between subimages to be correlated and thus prevent inaccuracy in phase estimation. Using an artificial test target mimicking the optical properties of a layer of living plant cells, an imaging performance with a Strehl ratio of approximately 0.5 was confirmed. Upon observation of chloroplast autofluorescence inside living leaf cells of the moss Physcomitrium patens, recovered resolution images were successfully obtained even with complex biological structures. Under bright-field illumination, the proposed method outperformed the conventional method, demonstrating the future potential of this method for label- and damage-free AO microscopy. Several points for improvement in terms of the effect of AO correction are discussed.

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

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 >0.7 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.

Deconvolution of partially compensated solar images from additional wavefront sensing.

A technique for restoring solar images partially compensated with adaptive optics is developed. An additional wavefront sensor is installed in an adaptive optics system to acquire residual wavefront information simultaneously to a solar image. A point spread function is derived from the wavefront information and used to deconvolve the solar image. Successful image restorations are demonstrated when the estimated point spread functions have relatively high Strehl ratios.

Application of self-deconvolution method to shift-and-add solar imaging.

A shift-and-add (SAA) operation is conducted to reconstruct a high-spatial-resolution image from atmospherically degraded solar images. The self-deconvolving data reconstruction algorithm is used to augment high-spatial-frequency components in solar speckle images and rectify the background component that results from the SAA operation. Self-deconvolved solar speckle images are shift and added and the resulting image shows high-spatial-resolution features.

Wind-flow measurement over the Subaru Telescope.

Wind flows over the 8.2-m Subaru Telescope at Mauna Kea in Hawaii were analyzed with a correlation method. Three or four wind flows were detected from our measurements. Spatial and temporal resolution of the wind-flow analysis across the 8.2 m pupil were investigated experimentally. A three-dimensional spatiotemporal-frequency analysis was also applied to the wind-flow data.

Blind deconvolution under band limitation.

A blind deconvolution problem is newly stated with the following conditions: the point-spread function is band limited, both the object and the point-spread function are nonnegative, and the solution is to be a diffraction-limited object. A blind deconvolution method was developed that can easily be applied to problems in optics because of the conditions used. The performance of the method is investigated with computer simulations.