Adaptive optics is rapidly transforming microscopy and high-resolution ophthalmic imaging. The adaptive elements commonly used to control optical wavefronts are liquid crystal spatial light modulators and deformable mirrors. We introduce a novel Multi-actuator Adaptive Lens that can correct aberrations to high order, and which has the potential to increase the spread of adaptive optics to many new applications by simplifying its integration with existing systems. Our method combines an adaptive lens with an imaged-based optimization control that allows the correction of images to the diffraction limit, and provides a reduction of hardware complexity with respect to existing state-of-the-art adaptive optics systems. The Multi-actuator Adaptive Lens design that we present can correct wavefront aberrations up to the 4th order of the Zernike polynomial characterization. The performance of the Multi-actuator Adaptive Lens is demonstrated in a wide field microscope, using a Shack-Hartmann wavefront sensor for closed loop control. The Multi-actuator Adaptive Lens and image-based wavefront-sensorless control were also integrated into the objective of a Fourier Domain Optical Coherence Tomography system for in vivo imaging of mouse retinal structures. The experimental results demonstrate that the insertion of the Multi-actuator Objective Lens can generate arbitrary wavefronts to correct aberrations down to the diffraction limit, and can be easily integrated into optical systems to improve the quality of aberrated images.

The mouse is an important model organism in ophthalmic research. As a part of the brain, the retina offers a unique opportunity to directly visualize changes associated with neurodegenerative disorders and vascular alterations, and to assess therapeutic interventions.¹ To better visualize the cellular microstructure and understand the molecular processes in the living retina, high-resolution retinal imaging of rodent eyes is desirable. Recently, two kinds of noninvasive retinal imaging modalities, scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT), have been successfully applied to monitor structural changes in the retinas of individual mice in longitudinal studies.¹ 2. 3. 4. 5. 6. 7. ^{– 8} SLO is a common diagnostic imaging technique in ophthalmology^9,10 and can generate images from retinal reflectance, autofluorescence, and extrinsic fluorescence. At the same time, the confocal arrangement helps SLO reject scattered light and improve image contrast, and potentially achieve moderate depth sectioning. In addition, an SLO can detect signals from exogenous fluorescence agents, such as fluorescein and indocyanine green, to improve imaging contrast of retinal and choroidal vasculature. Applied to mice, SLO also affords the possibility of imaging of cell-specific fluorescent proteins, introduced into the eye either through genetic engineering or by viral-mediated gene transfer.

Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a large-scale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.

The microsecond Er:YAG pulsed laser with a wavelength of λ = 2.94 μm has been widely used in the medical field, particularly for ablating dental tissues. Since bone and dental tissues have similar compositions, consisting of mineralized and rigid structures, the Er:YAG laser represents a promising tool for laserosteotomy applications. In this study, we explored the use of the Er:YAG laser for deep bone ablation, in an attempt to optimize its performance and identify its limitations. Tissue irrigation and the laser settings were optimized independently. We propose an automated irrigation feedback system capable of recognizing the temperature of the tissue and delivering water accordingly. The irrigation system used consists of a thin 50 μm diameter water jet. The water jet was able to penetrate deep into the crater during ablation, with a laminar flow length of 15 cm, ensuring the irrigation of deeper layers unreachable by conventional spray systems. Once the irrigation was optimized, ablation was considered independently of the irrigation water. In this way, we could better understand and adjust the laser parameters to suit our needs. We obtained line cuts as deep as 21 mm without causing any visible thermal damage to the surrounding tissue. The automated experimental setup proposed here has the potential to support deeper and faster ablation in laserosteotomy applications.