Monitoring Changes in Hemodynamics Following Optogenetic Stimulation

The brain is composed of billions of neurons, all of which connected through a vast network. After years of study and applications of different technologies and techniques, there are still more questions than answers when it comes to the fundamental functions of the brain. This project aims to provide a new tool which can be used to gain a better understanding of the fundamental mechanisms that govern neurological processes inside the brain. In order for neural networks to operate, blood has to be supplied through neighboring blood vessels. As such, the increase or decrease in the blood supply has been used as an indicator of neural activity. The neural activity and blood supply relationship is known as neural vasculature coupling. Monitoring the hemodynamics is used as an indicator of neurological activity, but the causal relationship is an area of current research. Gaining a better understanding of the coupling of neural activity and the surrounding vasculature provides a more accurate methodology to evaluate regional neural activity. The new optical technology applied in this project provides a set of tools to both stimulate and monitor this coupling relationship. Optogenetics provides the capability of stimulating neural activity using specific wavelengths of light. Essentially this tool allows for the direct stimulation of networks of neurons by simply shining one color of light onto the brain. Optical Coherence Tomography (OCT), another new optical technology applied in this project, can record volumetric images of blood vessels and flow using only infrared light. The combination of the two optical technologies is then capable of stimulating neural activity and monitoring the hemodynamic response inside the brain using only light. As a result of this project we have successfully demonstrated the capability of both stimulating and imaging the brain using new optical technologies. The optical stimulation of neural activity has evoked a direct hemodynamic effect as anticipated through neural-vasculature coupling. Changes in blood velocity, flow and dilatation were all recorded using the high resolution and high speed capabilities of the OCT system

Snapshot hyperspectral imaging of intracellular lasers

This work received financial support from a UK EPSRC Programme Grant (EP/P030017/1). PW was supported by the 1851 Research Fellowship from the Royal Commission. KD acknowledges support from the Australian Research Council (FL210100099). MCG acknowledges support from the Alexander von Humboldt Foundation (Humboldt professorship).Intracellular lasers are emerging as powerful biosensors for multiplexed tracking and precision sensing of cells and their microenvironment. This sensing capacity is enabled by quantifying their narrow-linewidth emission spectra, which is presently challenging to do at high speeds. In this work, we demonstrate rapid snapshot hyperspectral imaging of intracellular lasers. Using integral field mapping with a microlens array and a diffraction grating, we obtain images of the spatial and spectral intensity distribution from a single camera acquisition. We demonstrate widefield hyperspectral imaging over a 3 × 3 mm2 field of view and volumetric imaging over 250 × 250 × 800 µm3 (XYZ) volumes with a lateral (XY) resolution of 5 µm, axial (Z) resolution of 10 µm, and a spectral resolution of less than 0.8 nm. We evaluate the performance and outline the challenges and strengths of snapshot methods in the context of characterizing the emission from intracellular lasers. This method offers new opportunities for a diverse range of applications, including high-throughput and long-term biosensing with intracellular lasers.Publisher PDFPeer reviewe

COLOR MULTIPLEXED SINGLE PATTERN SLI

Structured light pattern projection techniques are well known methods of accurately capturing 3-Dimensional information of the target surface. Traditional structured light methods require several different patterns to recover the depth, without ambiguity or albedo sensitivity, and are corrupted by object movement during the projection/capture process. This thesis work presents and discusses a color multiplexed structured light technique for recovering object shape from a single image thus being insensitive to object motion. This method uses single pattern whose RGB channels are each encoded with a unique subpattern. The pattern is projected on to the target and the reflected image is captured using high resolution color digital camera. The image is then separated into individual color channels and analyzed for 3-D depth reconstruction through use of phase decoding and unwrapping algorithms thereby establishing the viability of the color multiplexed single pattern technique. Compared to traditional methods (like PMP, Laser Scan etc) only one image/one-shot measurement is required to obtain the 3-D depth information of the object, requires less expensive hardware and normalizes albedo sensitivity and surface color reflectance variations. A cosine manifold and a flat surface are measured with sufficient accuracy demonstrating the feasibility of a real-time system

No-scanning 3D measurement method using ultrafast dimensional conversion with a chirped optical frequency comb

A simultaneously high-precision, wide-range, and ultrafast time-resolution one-shot 3D shape measurement method is presented. Simultaneous times of flight from multiple positions to a target encoded in a chirped optical frequency comb can be obtained from spectral interferometry. We experimentally demonstrate a one-shot imaging profile measurement of a known step height of 480 µm with µm-level accuracy. We further demonstrate the extension of the dynamic range by measuring in one shot a large step height of 3 m while maintaining high accuracy using the accurate pulse-to-pulse separation of the optical frequency comb. The proposed method with its large dynamic range and measurement versatility can be applied to a broad range of applications, including microscopic structures, objects with large size or aspect ratio, and ultrafast time-resolved imaging. This study provides a powerful and versatile tool for 3D measurement, where various ranges of measurement performances can be tailored to demand

Optogenetic Interrogation and Manipulation of Vascular Blood Flow in Cortex

Understanding blood flow regulatory mechanisms that correlate the regional blood flow with the level of local neuronal activity in brain is an ongoing research. Discerning different aspects of this coupling is of substantial importance in interpretation of functional imaging results, such as functional magnetic resonance imaging (fMRI), that rely on hemodynamic recordings to detect and image brain neuronal activity. Moreover, this understanding can provide insight into blood flow disorders under different pathophysiological conditions and possible treatments for such disorders. The blood regulatory mechanisms can be studied at two different; however, complementary levels: at the cellular level or at the vascular level. To fully understand the regulatory mechanisms in brain, it is essential to discern details of the coupling mechanism in each level. While, the cellular pathways of the coupling mechanism has been studied extensively in the past few decades, our understanding of the vascular response to brain activity is fairly basic. The main objective of this dissertation is to develop proper methods and instrumentation to interrogate regional cortical vasodynamics in response to local brain stimulation. For this purpose we offer the design of a custom-made OCT scanner and the necessary lens mechanisms to integrate the OCT system, fluorescence imaging, and optogenetic stimulation technologies in a single system. The design uses off-the-shelf components for a cost-effective design. The modular design of the device allows scientists to modify it in accordance with their research needs. With this multi-modal system we are able to monitor blood flow, blood velocity, and lumen diameter of pial vessels, simultaneously. Additionally, the system design provides the possibility of generating arbitrary spatial stimulation light pattern on brain. These abilities enables researchers to capture more diverse datasets and, eventually, obtain a more comprehensive picture of the vasodynamics in the brain. Along with the device we also proposed new biological experiments that are tailored to investigate the spatio-temporal properties of the vascular response to optical neurostimulation of the excitatory neurons. We demonstrate the ability of the proposed methods to investigate the effect of length and amplitude of stimulation on the temporal pattern of response in the blood flow, blood velocity, and diameter of the pial vessels. Moreover, we offer systemic approaches to investigate the spatial characteristics of the response in a vascular network. In these methods we apply arbitrary spatial patterns of optical stimulation to the cortex of transgenic mice and monitor the attributes of surrounding vessels. With this flexibility we were able to image the brain region that is influenced by a pial artery. After characterizing the spatio-temporal properties of the vascular blood flow response to optical neuro-modulation, we demonstrate the design and application of an optogenetic-based closed-loop controller mechanism in the brain. This controller, uses a proportional–integral–derivative (PID) compensator to engineer temporal optogenetic stimulation light pulses and maintain the flow of blood at various user defined levels in a set of selected arteries. Upon tuning the gain values of the PID controller we obtained a near to critically-damped response in the blood flow of selected arterial vessels

Optical MEMS

Optical microelectromechanical systems (MEMS), microoptoelectromechanical systems (MOEMS), or optical microsystems are devices or systems that interact with light through actuation or sensing at a micro- or millimeter scale. Optical MEMS have had enormous commercial success in projectors, displays, and fiberoptic communications. The best-known example is Texas Instruments’ digital micromirror devices (DMDs). The development of optical MEMS was impeded seriously by the Telecom Bubble in 2000. Fortunately, DMDs grew their market size even in that economy downturn. Meanwhile, in the last one and half decade, the optical MEMS market has been slowly but steadily recovering. During this time, the major technological change was the shift of thin-film polysilicon microstructures to single-crystal–silicon microsructures. Especially in the last few years, cloud data centers are demanding large-port optical cross connects (OXCs) and autonomous driving looks for miniature LiDAR, and virtual reality/augmented reality (VR/AR) demands tiny optical scanners. This is a new wave of opportunities for optical MEMS. Furthermore, several research institutes around the world have been developing MOEMS devices for extreme applications (very fine tailoring of light beam in terms of phase, intensity, or wavelength) and/or extreme environments (vacuum, cryogenic temperatures) for many years. Accordingly, this Special Issue seeks to showcase research papers, short communications, and review articles that focus on (1) novel design, fabrication, control, and modeling of optical MEMS devices based on all kinds of actuation/sensing mechanisms; and (2) new developments of applying optical MEMS devices of any kind in consumer electronics, optical communications, industry, biology, medicine, agriculture, physics, astronomy, space, or defense