Various Applications of Methods and Elements of Adaptive Optics

This volume is focused on a wide range of topics, including adaptive optic components and tools, wavefront sensing, different control algorithms, astronomy, and propagation through turbulent and turbid media

The development of a high speed 3D 2-photon microscope for neuroscience

The progress of neuroscience is limited by the instrumentation available to it for studying the brain. At present, there is a serious instrumentation gap between functional Magnetic Resonance Imaging (fMRI) of whole brains and the microscopic scale functional imaging possible with today’s optical microscopes and electrophysiology techniques, such as patch clamping of individual neurons. This thesis describes the development of a new extension to optical microscopy that enables refocusing within 25 microseconds rather than the large fraction of a second possible by moving the sample or objective. The system is capable of refocusing a laser beam that is monitoring activity in 3D samples of live brain tissue 300 times faster than previously possible. This will make practical a new type of optical functional imaging for studying small sub-networks of neurons containing up to about 30,000 neurons at up to 30,000 sub micrometre sized monitored points of interest per second. The thesis describes the development of a detailed design for a new type of 3D scanner that uses Acousto-Optic Deflectors (AODs) to diffractively deflect and focus an intense laser beam beneath a conventional microscope objective. The fluorescence of calcium sensitive dyes in live neurons is used to monitor action potentials conveying signals between neurons. The optical and systems engineering problems and design trade-offs involved are discussed in detail. The results of extensive computer modelling are described and innovative solutions to several key optical physics based engineering problems are explained. The practical problems found in building a prototype machine incorporating these innovations are described and the encouraging first operational results from the machine reported

Astigmatism and Pseudoaccommodation in Pseudophakic Eyes

noAdvanced IOLs with circumferential zones of different power provide pseudoaccommodation. We investigated the potential for power variation with meridian, namely astigmatism, to provide pseudo-accommodation. With appropriate power and axis orientations, acceptable pseudo-accommodation can be achieved

Image Reconstruction in Photoacoustic Computed Tomography with Acoustically Heterogeneous Media

Photoacoustic computed tomography (PACT), also known as optoacoustic or thermoacoustic tomography, is a rapidly emerging hybrid imaging modality that combines optical image contrast with ultrasound detection. The majority of currently available PACT image reconstruction algorithms are based on idealized imaging models that assume a lossless and acoustically homogeneous medium. However, in many applications of PACT these assumptions are violated and the induced photoacoustic (PA) wavefields are scattered and absorbed as they propagate to the receiving transducers. In those applications of PACT, the reconstructed images can contain significant distortions and artifacts if the inhomogeneous acoustic properties of the object are not accounted for in the reconstruction algorithm. In this dissertation, we develop and investigate a full-wave approach to iterative image reconstruction in PACT with acoustically heterogeneous lossy media. A key contribution of this work is the establishment of a discrete imaging model that is based on the exact PA wave equation and a procedure to implement an associated matched discrete forward and backprojection operator pair, which permits application of a variety of modern iterative image reconstruction algorithms that can mitigate the effects of noise, data incompleteness and model errors. Another key contribution is the development of an optimization approach to joint reconstruction (JR) of absorbed optical energy density and speed of sound in PACT, which is utilized to investigate the numerical properties of the JR problem and its feasibility in practice. We also develop a TR-based methodology to compensate for heterogeneous acoustic attenuation that obeys a frequency power law. In addition, we propose a image reconstruction methodology for transcranial PACT that employs detailed subject-specific descriptions of the acoustic properties of the skull to mitigate skull-induced distortions in the reconstructed image

Ultrasound Imaging

This book provides an overview of ultrafast ultrasound imaging, 3D high-quality ultrasonic imaging, correction of phase aberrations in medical ultrasound images, etc. Several interesting medical and clinical applications areas are also discussed in the book, like the use of three dimensional ultrasound imaging in evaluation of Asherman's syndrome, the role of 3D ultrasound in assessment of endometrial receptivity and follicular vascularity to predict the quality oocyte, ultrasound imaging in vascular diseases and the fetal palate, clinical application of ultrasound molecular imaging, Doppler abdominal ultrasound in small animals and so on

High-sensitivity interferometry

High-sensitivity interferometric techniques are considered for non-destructive testing applications. The methods enable quantitative measurement of optical path variations, resulting from dynamic changes within the test object. [Continues.

Fast Neuronal Imaging using Objective Coupled Planar Illumination Microscopy

Complex computations performed by the brain are produced by activities of neuronal populations. There is a large diversity in the functions of each individual neuron, and neuronal activities occur in the time scale of milliseconds. In order to gain a fundamental understanding of the neuronal populations, one has to measure activity of each neuron at high temporal resolution, while investigating enough neurons to encapsulate the neuronal diversity. Traditional neurotechniques such as electrophysiology and optical imaging are constrained by the number of neurons whose activities can be simultaneously measured or the speed of measuring such activities. We have developed a novel light-sheet based technique called Objective Coupled Planar Illumination: OCPI) microscopy which is capable of measuring simultaneous activities of thousands of neurons at high speeds. In this thesis I pursue the following two aims: * Improve OCPI microscopy by enhancing the spatial resolution deeper in tissue. Tissue inhomogeneity and refractive index mismatch at the surface of the tissue lead to optical aberrations. We have compensated for such aberrations by: 1) miniaturizing the OCPI illumination optics, so as to enable more vertical imaging of the tissue,: 2) correcting for the angular defocus caused by the refraction at the immersion fluid/tissue interface, and: 3) applying adaptive optics to correct for higher order optical aberrations. The improvement in the depth at which one can image tissue will enable the measurement of activities of neuronal populations in cortical areas. * Measure the diversity in the expression pattern of VSNs responsive to sulfated steroids. Nodari et al. have identified sulfated steroids as a novel family of ligands which activate vomeronasal sensory neurons: VSNs). Due to the experimental constraints, it has not been possible to obtain a comprehensive understanding of the number, location and functional characteristics of the sulfated steroid responsive VSNs. Applying OCPI microscopy and calcium imaging to simultaneously image thousands of VSNs, we show that the sulfated steroid responsive neurons: 1) have unique ligand preferences,: 2) are predominantly present in the apical regions of the VNO, and: 3) that the choice of expression of a receptor type is not purely stochastic

Developing Wavefront Shaping Techniques for Focusing through Highly Dynamic Scattering Media

One of the prime limiting factors of optical imaging in biological applications is the diffusion of light by tissue, which prevents focusing at depths greater than the optical diffusion limit of ~1 mm in soft tissue. This greatly restricts the utility of optical diagnostic and therapeutic techniques, such as optogenetics, microsurgery, optical tweezing, and phototherapy of deep tissue, which require focused light in order to function. Wavefront shaping extends the depth at which optical focusing may be achieved by compensating for phase distortions induced by scattering, allowing for focusing through constructive interference. However, due to physiological motion, scattering of light in tissue is deterministic only within a brief speckle correlation time. In in vivo soft tissue, this speckle correlation is on the order of milliseconds. Because wavefront shaping relies on deterministic scattering in order to compensate for the resulting phase distortion, the wavefront must be optimized within this brief period. This presents a challenge as the speed of digital wavefront shaping has typically been limited by the relatively long time required to measure and display the optimal phase pattern due to the low speed of cameras, data transfer and processing, and spatial light modulators. In order to overcome these restrictions, wavefront shaping techniques which minimize the time required in measurement and display are therefore vital. In this dissertation, I will describe our efforts to improve the speed of wavefront shaping without sacrificing the performance of the systems. To this end, we have successfully developed several systems which are capable of full-phase wavefront shaping with latencies of 9 ms or less. In addition, we report an all-digital alignment compensation protocol, which may be used to obtain optimal alignment in digital optical phase conjugation systems, a key component when acquiring the best possible focusing performance