Adaptive Optics (AO) has revealed as a very promising technique for high-resolution microscopy, where the presence of optical aberrations can easily compromise the image quality. Typical AO systems however, are almost impossible to implement on commercial microscopes. We propose a simple approach by using a Multi-actuator Adaptive Lens (MAL) that can be inserted right after the objective and works in conjunction with an image optimization software allowing for a wavefront sensorless correction. We presented the results obtained on several commercial microscopes among which a confocal microscope, a fluorescence microscope, a light sheet microscope and a multiphoton microscope.
It has been widely proven in literature that most optical microscopy techniques can greatly benefit from the application of adaptive optics correction of phase aberrations through an adaptive optical element, such as a deformable mirror or a spatial light modulator. However, adaptive optics is not yet widely adopted in the life sciences community, mostly due to the lack of adaptive commercial microscopy systems, and the inherent technical difficulty in modifying an existing microscopy setup to integrate an adaptive element, both on the software and hardware sides. We present a plug-and-play adaptive optics module for generic optical microscopes, based on a prototype refractive 18 actuators adaptive optical element, which can be inserted in any microscope between the objective and the microscope body. Correction is performed in a sensorless fashion, optimizing image quality metrics of the image presented to the user on screen. The results presented show how an end-user oriented commercial confocal laser scanning microscope (Leica SP5) can be upgraded with adaptive optics with minor hardware modifications, and no changes to the microscope control software.
Optical Coherence Tomography (OCT) has revolutionized modern ophthalmology, providing depth resolved images of the retinal layers in a system that is suited to a clinical environment. A limitation of the performance and utilization of the OCT systems has been the lateral resolution. Through the combination of wavefront sensorless adaptive optics with dual variable optical elements, we present a compact lens based OCT system that is capable of imaging the photoreceptor mosaic. We utilized a commercially available variable focal length lens to correct for a wide range of defocus commonly found in patient eyes, and a multi-actuator adaptive lens after linearization of the hysteresis in the piezoelectric actuators for aberration correction to obtain near diffraction limited imaging at the retina. A parallel processing computational platform permitted real-time image acquisition and display. The Data-based Online Nonlinear Extremum seeker (DONE) algorithm was used for real time optimization of the wavefront sensorless adaptive optics OCT, and the performance was compared with a coordinate search algorithm. Cross sectional images of the retinal layers and en face images of the cone photoreceptor mosaic acquired in vivo from research volunteers before and after WSAO optimization are presented. Applying the DONE algorithm in vivo for wavefront sensorless AO-OCT demonstrates that the DONE algorithm succeeds in drastically improving the signal while achieving a computational time of 1 ms per iteration, making it applicable for high speed real time applications.
Adaptive optics has been successfully applied to cellular resolution imaging of the retina, enabling visualization of the characteristic mosaic patterns of the outer retina. Wavefront sensorless adaptive optics (WSAO) is a novel technique that facilitates high resolution ophthalmic imaging; it replaces the Hartmann-Shack Wavefront Sensor with an image-driven optimization algorithm and mitigates some the challenges encountered with sensor-based designs. However, WSAO generally requires longer time to perform aberrations correction than the conventional closed-loop adaptive optics. When used for in vivo retinal imaging applications, motion artifacts during the WSAO optimization process will affect the quality of the aberration correction. A faster converging optimization scheme needs to be developed to account for rapid temporal variation of the wavefront and continuously apply corrections. In this project, we investigate the Databased Online Nonlinear Extremum-seeker (DONE), a novel non-linear multivariate optimization algorithm in combination with in vivo human WSAO OCT imaging. We also report both hardware and software updates of our compact lens based WSAO 1060nm swept source OCT human retinal imaging system, including real time retinal layer segmentation and tracking (ILM and RPE), hysteresis correction for the multi-actuator adaptive lens, precise synchronization control for the 200kHz laser source, and a zoom lens unit for rapid switching of the field of view. Cross sectional images of the retinal layers and en face images of the cone photoreceptor mosaic acquired in vivo from research volunteers before and after WSAO optimization are presented.
The use of adaptive lenses instead of deformable mirrors can simplify the implementation of an adaptive optics system. The recently introduced Multi-actuator Adaptive Lens (MAL) can be used in closed loop with a wavefront sensor to correct for time-variant wavefront aberrations. The MAL can guarantee a level of correction and a response time similar to the ones obtained with deformable mirrors. The adaptive lens is based on the use of piezoelectric actuators and, without any obstruction or electrodes in the clear aperture, can guarantee a fast response time, less than ~10ms. Our tests show that the MAL can be used both in combination with a wavefront sensor in a “classical” adaptive optics closed loop, or in a wavefront sensorless configuration. The latter has allowed us to design more compact and simple imaging systems for different microscopy platforms. We will show that the Multi-actuator Adaptive Lens has been successfully used for in-vivo OCT ophthalmic imaging in both mice and humans, as well as confocal and two photon microscopy. We tested and compared different optimization strategies such as coordinate search and the DONE algorithm. The results suggest that the MAL optimization can correct for eye aberrations with a pupil of 5mm or sample induced aberrations in microscopy.
In this manuscript, we present a lens setup for large defocus and astigmatism correction. A deformable defocus lens and two rotational cylindrical lenses are used to control the defocus and astigmatism. The setup is calibrated using a simple model that allows the calculation of the lens inputs so that a desired defocus and astigmatism are actuated on the eye. The setup is tested by determining the feedforward prediction error, imaging a resolution target, and removing introduced aberrations.
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