Physical Sciences and Mathematics Commons^™

6d Single-Fluorogen Orientation-Localization Microscopy For Elucidating The Architecture Of Beta-Sheet Assemblies And Biomolecular Condensates, Tingting Wu, Weiyan Zhou, Jai S. Rudra, Rohit V. Pappu, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

We develop six-dimensional single-molecule orientation-localization microscopy (SMOLM) to measure the 3D positions and 3D orientations simultaneously of single fluorophores. We show how careful optimization of phase and polarization modulation components can encode phase, polarization, and angular spectrum information from each fluorescence photon into a microscope’s dipole-spread function. We used the transient binding and blinking of Nile red (NR) to characterize the helical structure of fibrils formed by designed amphipathic peptides, KFE8^L and KFE8^D, and the pathological amyloid-beta peptide Aβ42. We also deployed merocyanine 540 to uncover the interfacial architectures of biomolecular condensates.

Six-Dimensional Single-Molecule Imaging With Isotropic Resolution Using A Multi-View Reflector Microscope, Oumeng Zhang, Zijian Guo, Yuanyuan He, Tingting Wu, Michael D. Vahey, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Imaging of both the positions and orientations of single fluorophores, termed single-molecule orientation-localization microscopy, is a powerful tool for the study of biochemical processes. However, the limited photon budget associated with single-molecule fluorescence makes high-dimensional imaging with isotropic, nanoscale spatial resolution a formidable challenge. Here we realize a radially and azimuthally polarized multi-view reflector (raMVR) microscope for the imaging of the three-dimensional (3D) positions and 3D orientations of single molecules, with precisions of 10.9 nm and 2.0° over a 1.5-μm depth range. The raMVR microscope achieves 6D super-resolution imaging of Nile red molecules transiently bound to lipid-coated spheres, accurately resolving …

Model-Based Deep Learning For Computational Imaging, Xiaojian Xu

McKelvey School of Engineering Theses & Dissertations

This dissertation addresses model-based deep learning for computational imaging. The motivation of our work is driven by the increasing interests in the combination of imaging model, which provides data-consistency guarantees to the observed measurements, and deep learning, which provides advanced prior modeling driven by data. Following this idea, we develop multiple algorithms by integrating the classical model-based optimization and modern deep learning to enable efficient and reliable imaging. We demonstrate the performance of our algorithms by validating their performance on various imaging applications and providing rigorous theoretical analysis.

The dissertation evaluates and extends three general frameworks, plug-and-play priors (PnP), regularized …

Design & Analysis Of Mixed-Mode Integrated Circuit For Pulse-Shape Discrimination, Bryan Orabutt

McKelvey School of Engineering Theses & Dissertations

In nuclear science experiments it is usually necessary to determine the type of radiation, its energy and direction with considerable accuracy. The detection of neutrons and discriminating them from gamma rays is particularly difficult. A popular method of doing so is to measure characteristics intrinsic to the pulse shape of each radiation type in order to perform pulse-shape discrimination (PSD).

Historically, PSD capable systems have been designed with two approaches in mind: specialized analog circuitry, or digital signal processing (DSP). In this work we propose a PSD capable circuit topology using techniques from both the analog and DSP domains. We …

Resolving The Three-Dimensional Rotational And Translational Dynamics Of Single Molecules Using Radially And Azimuthally Polarized Fluorescence, Oumeng Zhang, Weiyan Zhou, Jin Lu, Tingting Wu, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

We report a radially and azimuthally polarized (raPol) microscope for high detection and estimation performance in single-molecule orientation-localization microscopy (SMOLM). With 5000 photons detected from Nile red (NR) transiently bound within supported lipid bilayers (SLBs), raPol SMOLM achieves 2.9 nm localization precision, 1.5° orientation precision, and 0.17 sr precision in estimating rotational wobble. Within DPPC SLBs, SMOLM imaging reveals the existence of randomly oriented binding pockets that prevent NR from freely exploring all orientations. Treating the SLBs with cholesterol-loaded methyl-β-cyclodextrin (MβCD-chol) causes NR’s orientational diffusion to be dramatically reduced, but curiously NR’s median lateral displacements drastically increase from 20.8 to …

Single-Molecule Localization Microscopy Of 3d Orientation And Anisotropic Wobble Using A Polarized Vortex Point Spread Function, Tianben Ding, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Within condensed matter, single fluorophores are sensitive probes of their chemical environments, but it is difficult to use their limited photon budget to image precisely their positions, 3D orientations, and rotational diffusion simultaneously. We demonstrate the polarized vortex point spread function (PSF) for measuring these parameters, including characterizing the anisotropy of a molecule’s wobble, simultaneously from a single image. Even when imaging dim emitters (∼500 photons detected), the polarized vortex PSF can obtain 12 nm localization precision, 4°–8° orientation precision, and 26° wobble precision. We use the vortex PSF to measure the emission anisotropy of fluorescent beads, the wobble dynamics …

Machine Learning For Analog/Mixed-Signal Integrated Circuit Design Automation, Weidong Cao

McKelvey School of Engineering Theses & Dissertations

Analog/mixed-signal (AMS) integrated circuits (ICs) play an essential role in electronic systems by processing analog signals and performing data conversion to bridge the analog physical world and our digital information world.Their ubiquitousness powers diverse applications ranging from smart devices and autonomous cars to crucial infrastructures. Despite such critical importance, conventional design strategies of AMS circuits still follow an expensive and time-consuming manual process and are unable to meet the exponentially-growing productivity demands from industry and satisfy the rapidly-changing design specifications from many emerging applications. Design automation of AMS IC is thus the key to tackling these challenges and has been …

Continuous-Time And Complex Growth Transforms For Analog Computing And Optimization, Oindrila Chatterjee

McKelvey School of Engineering Theses & Dissertations

Analog computing is a promising and practical candidate for solving complex computational problems involving algebraic and differential equations. At the fundamental level, an analog computing framework can be viewed as a dynamical system that evolves following fundamental physical principles, like energy minimization, to solve a computing task. Additionally, conservation laws, such as conservation of charge, energy, or mass, provide a natural way to couple and constrain spatially separated variables. Taking a cue from these observations, in this dissertation, I have explored a novel dynamical system-based computing framework that exploits naturally occurring analog conservation constraints to solve a variety of optimization …

Algebraic, Computational, And Data-Driven Methods For Control-Theoretic Analysis And Learning Of Ensemble Systems, Wei Miao

McKelvey School of Engineering Theses & Dissertations

In this thesis, we study a class of problems involving a population of dynamical systems under a common control signal, namely, ensemble systems, through both control-theoretic and data-driven perspectives. These problems are stemmed from the growing need to understand and manipulate large collections of dynamical systems in emerging scientific areas such as quantum control, neuroscience, and magnetic resonance imaging. We examine fundamental control-theoretic properties such as ensemble controllability of ensemble systems and ensemble reachability of ensemble states, and propose ensemble control design approaches to devise control signals that steer ensemble systems to desired profiles. We show that these control-theoretic properties …

Aerosol Vapor Synthesis Of Organic Processable Pedot Particles And Measuring Electric Conductivity Using A 3d Printed Probe Station, Yang Lu

McKelvey School of Engineering Theses & Dissertations

Conducting polymers are organic semiconductors characterized by conjugated backbones (alternating single-double bonds) that enable mixed ionic-electronic conductivity. Their polymeric nature, tunable band structure and reversible redox capability have demonstrated fundamental advances in the fields ranging from electrochemical energy storage, sensing, to electro/photo catalysis and neuromorphic engineering. Conjugated backbones, the origin of all the unique physical and chemical properties associated with conducting polymers, prevent their solubility due to high lattice energy which hinders processing. Current solution utilizes a long-chain polymer (PSS) as dopants to render conducting polymer water dispersible (PEDOT:PSS). Nonetheless, PSS is highly acidic and hydrophilic limiting applicability with acid-incompatible …

Computational Modelling Enables Robust Multidimensional Nanoscopy, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

The following sections are included:

• Present State of Computational Modelling in Fluorescence Nanoscopy

• Recent Contributions to Computational Modelling in Fluorescence Nanoscopy

• Outlook on Computational Modelling in Fluorescence Nanoscopy

• Acknowledgments

• References

Deep Learning For Task-Based Image Quality Assessment In Medical Imaging, Weimin Zhou

McKelvey School of Engineering Theses & Dissertations

It has been advocated to use objective measures of image quality (IQ) for assessing and optimizing medical imaging systems. Objective measures of IQ quantify the performance of an observer at a specific diagnostic task. Binary signal detection tasks and joint signal detection and localization (detection-localization) tasks are commonly considered in medical imaging. When optimizing imaging systems for binary signal detection tasks, the performance of the Bayesian Ideal Observer (IO) has been advocated for use as a figure-of-merit (FOM). The IO maximizes the observer performance that is summarized by the receiver operating characteristic (ROC) curve. When signal detection-localization tasks are considered, …

Holistic Control For Cyber-Physical Systems, Yehan Ma

McKelvey School of Engineering Theses & Dissertations

The Industrial Internet of Things (IIoT) are transforming industries through emerging technologies such as wireless networks, edge computing, and machine learning. However, IIoT technologies are not ready for control systems for industrial automation that demands control performance of physical processes, resiliency to both cyber and physical disturbances, and energy efficiency. To meet the challenges of IIoT-driven control, we propose holistic control as a cyber-physical system (CPS) approach to next-generation industrial automation systems. In contrast to traditional industrial automation systems where computing, communication, and control are managed in isolation, holistic control orchestrates the management of cyber platforms (networks and computing platforms) …

Machine Learning Morphisms: A Framework For Designing And Analyzing Machine Learning Work Ows, Applied To Separability, Error Bounds, And 30-Day Hospital Readmissions, Eric Zenon Cawi

McKelvey School of Engineering Theses & Dissertations

A machine learning workflow is the sequence of tasks necessary to implement a machine learning application, including data collection, preprocessing, feature engineering, exploratory analysis, and model training/selection. In this dissertation we propose the Machine Learning Morphism (MLM) as a mathematical framework to describe the tasks in a workflow. The MLM is a tuple consisting of: Input Space, Output Space, Learning Morphism, Parameter Prior, Empirical Risk Function. This contains the information necessary to learn the parameters of the learning morphism, which represents a workflow task. In chapter 1, we give a short review of typical tasks present in a workflow, as …

Single‐Molecule 3d Orientation Imaging Reveals Nanoscale Compositional Heterogeneity In Lipid Membranes, Jin Lu, Hesam Mazidi, Tianben Ding, Oumeng Zhang, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

In soft matter, thermal energy causes molecules to continuously translate and rotate, even in crowded environments, thereby impacting the spatial organization and function of most molecular assemblies, such as lipid membranes. Directly measuring the orientation and spatial organization of large collections (>3000 molecules μm⁻²) of single molecules with nanoscale resolution remains elusive. In this paper, we utilize SMOLM, single‐molecule orientation localization microscopy, to directly measure the orientation spectra (3D orientation plus “wobble”) of lipophilic probes transiently bound to lipid membranes, revealing that Nile red's (NR) orientation spectra are extremely sensitive to membrane chemical composition. SMOLM images resolve …

Structural Organization And Chemical Activity Revealed By New Developments In Single-Molecule Fluorescence And Orientation Imaging, Tianben Ding

McKelvey School of Engineering Theses & Dissertations

Single-molecule (SM) fluorescence and its localization are important and versatile tools for understanding and quantifying dynamical nanoscale behavior of nanoparticles and biological systems. By actively controlling the concentration of fluorescent molecules and precisely localizing individual single molecules, it is possible to overcome the classical diffraction limit and achieve 'super-resolution' with image resolution on the order of 10 nanometers.

Single molecules also can be considered as nanoscale sensors since their fluorescence changes in response to their local nanoenvironment. This dissertation discusses extending this SM approach to resolve heterogeneity and dynamics of nanoscale materials and biophysical structures by using positions and orientations …

Convex Relaxations For Particle-Gradient Flow With Applications In Super-Resolution Single-Molecule Localization Microscopy, Hesam Mazidisharfabadi

McKelvey School of Engineering Theses & Dissertations

Single-molecule localization microscopy (SMLM) techniques have become advanced bioanalytical tools by quantifying the positions and orientations of molecules in space and time at the nanoscale. With the noisy and heterogeneous nature of SMLM datasets in mind, we discuss leveraging particle-gradient flow 1) for quantifying the accuracy of localization algorithms with and without ground truth and 2) as a basis for novel, model-driven localization algorithms with empirically robust performance. Using experimental data, we demonstrate that overlapping images of molecules, a typical consequence of densely packed biological structures, cause biases in position estimates and reconstruction artifacts. To minimize such biases, we develop …

Investigating Single Precision Floating General Matrix Multiply In Heterogeneous Hardware, Steven Harris

McKelvey School of Engineering Theses & Dissertations

The fundamental operation of matrix multiplication is ubiquitous across a myriad of disciplines. Yet, the identification of new optimizations for matrix multiplication remains relevant for emerging hardware architectures and heterogeneous systems. Frameworks such as OpenCL enable computation orchestration on existing systems, and its availability using the Intel High Level Synthesis compiler allows users to architect new designs for reconfigurable hardware using C/C++. Using the HARPv2 as a vehicle for exploration, we investigate the utility of several of the most notable matrix multiplication optimizations to better understand the performance portability of OpenCL and the implications for such optimizations on this and …

Measuring Localization Confidence For Quantifying Accuracy And Heterogeneity In Single-Molecule Super-Resolution Microscopy, Hesam Mazidi, Tianben Ding, Arye Nehorai, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

We present a computational method, termed Wasserstein-induced flux (WIF), to robustly quantify the accuracy of individual localizations within a single-molecule localization microscopy (SMLM) dataset without ground- truth knowledge of the sample. WIF relies on the observation that accurate localizations are stable with respect to an arbitrary computational perturbation. Inspired by optimal transport theory, we measure the stability of individual localizations and develop an efficient optimization algorithm to compute WIF. We demonstrate the advantage of WIF in accurately quantifying imaging artifacts in high-density reconstruction of a tubulin network. WIF represents an advance in quantifying systematic errors with unknown and complex distributions, …

A Computationally-Efficient Bound For The Variance Of Measuring The Orientation Of Single Molecules, Tingting Wu, Tianben Ding, Hesam Mazidi, Oumeng Zhang, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Modulating the polarization of excitation light, resolving the polarization of emitted fluorescence, and point spread function (PSF) engineering have been widely leveraged for measuring the orientation of single molecules. Typically, the performance of these techniques is optimized and quantified using the Cramér-Rao bound (CRB), which describes the best possible measurement variance of an unbiased estimator. However, CRB is a local measure and requires exhaustive sampling across the measurement space to fully characterize measurement precision. We develop a global variance upper bound (VUB) for fast quantification and comparison of orientation measurement techniques. Our VUB tightly bounds the diagonal elements of the …

Instrumentation For Dynamic Nuclear Polarization And Application Of Electron Decoupling For Electron Relaxation Measurement, Nicholas Howard Alaniva

Arts & Sciences Electronic Theses and Dissertations

Dynamic nuclear polarization nuclear magnetic resonance (DNP NMR) exploits internal electron spin and nuclear spin interactions to increase sensitivity and uncover valuable information regarding structure and dynamics of a system. To manipulate these interactions, instrumentation is developed to combine high-power microwave and radiofrequency irradiation with the ability to spin samples at the magic angle (MAS) at temperatures from 90 K to 4.2 K. Electron decoupling uses frequency-modulated microwaves to mitigate the electron-nuclear dipolar interaction, improving signal intensity and resolution in DNP NMR experiments. Electron decoupling is combined with short DNP periods to encode electron spin information in polarized nuclear signal. …

Polarization Division Multiplexing For Optical Data Communications, Darko Ivanovich

McKelvey School of Engineering Theses & Dissertations

Multiple parallel channels are ubiquitous in optical communications, with spatial division multiplexing (separate physical paths) and wavelength division multiplexing (separate optical wavelengths) being the most common forms. In this research work, we investigate the viability of polarization division multiplexing, the separation of distinct parallel optical communication channels through the polarization properties of light. We investigate polarization division multiplexing based optical communication systems in five distinct parts. In the first part of the work, we define a simulation model of two or more linearly polarized optical signals (at different polarization angles) that are transmitted through a common medium (e.g., air), filtered …

Erratum: "Imaging The Three‐Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri‐Spot Point Spread Function", Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

In the original paper, a calibration error exists in the image-formation model used to analyze experimental images taken by our microscope, causing a bias in the orientation measurements in Figs. 2 and 3. The updated measurements are shown in Fig. E1. We have also updated the supplementary material for the original article to discuss the revised PSF model and estimation algorithms (supplementary material 2) and show the revised model and measurements (Figs. S1, S3, S7, S8, and S10–S13).

Fundamental Limits Of Measuring Single-Molecule Rotational Mobility, Oumeng Zhang, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Various methods exist for measuring molecular orientation, thereby providing insight into biochemical activities at nanoscale. Since fluorescence intensity and not electric field is detected, these methods are limited to measuring even-order moments of molecular orientation. However, any measurement noise, for example photon shot noise, will result in nonzero measurements of any of these even-order moments, thereby causing rotationally-free molecules to appear to be partially constrained. Here, we build a model to quantify measurement errors in rotational mobility. Our theoretical framework enables scientists to choose the optimal single-molecule orientation measurement technique for any desired measurement accuracy and photon budget.

Nanopower Analog Frontends For Cyber-Physical Systems, Kenji Aono

McKelvey School of Engineering Theses & Dissertations

In a world that is increasingly dominated by advances made in digital systems, this work will explore the exploiting of naturally occurring physical phenomena to pave the way towards a self-powered sensor for Cyber-Physical Systems (CPS). In general, a sensor frontend can be broken up into a handful of basic stages: transduction, filtering, energy conversion, measurement, and interfacing. One analog artifact that was investigated for filtering was the physical phenomenon of hysteresis induced in current-mode biquads driven near or at their saturation limit. Known as jump resonance, this analog construct facilitates a higher quality factor to be brought about without …

Self-Powered Time-Keeping And Time-Of-Occurrence Sensing, Liang Zhou

McKelvey School of Engineering Theses & Dissertations

Self-powered and passive Internet-of-Things (IoT) devices (e.g. RFID tags, financial assets, wireless sensors and surface-mount devices) have been widely deployed in our everyday and industrial applications. While diverse functionalities have been implemented in passive systems, the lack of a reference clock limits the design space of such devices used for applications such as time-stamping sensing, recording and dynamic authentication. Self-powered time-keeping in passive systems has been challenging because they do not have access to continuous power sources. While energy transducers can harvest power from ambient environment, the intermittent power cannot support continuous operation for reference clocks. The thesis of this …

Robust Engineering Of Dynamic Structures In Complex Networks, Walter Botongo Bomela

McKelvey School of Engineering Theses & Dissertations

Populations of nearly identical dynamical systems are ubiquitous in natural and engineered systems, in which each unit plays a crucial role in determining the functioning of the ensemble. Robust and optimal control of such large collections of dynamical units remains a grand challenge, especially, when these units interact and form a complex network. Motivated by compelling practical problems in power systems, neural engineering and quantum control, where individual units often have to work in tandem to achieve a desired dynamic behavior, e.g., maintaining synchronization of generators in a power grid or conveying information in a neuronal network; in this dissertation, …

Instrumentation For Cryogenic Dynamic Nuclear Polarization And Electron Decoupling In Rotating Solids, Faith Joellen Scott

Arts & Sciences Electronic Theses and Dissertations

Dynamic nuclear polarization (DNP) increases the sensitivity of nuclear magnetic resonance (NMR) using the higher polarization of electron radical spins compared to nuclear spins. The addition of electron radicals for DNP to the sample can cause hyperfine broadening, which decreases the resolution of the NMR resonances due to hyperfine interactions between electron and nuclear spins. Electron decoupling has been shown to attenuate the effects of hyperfine coupling in rotating solids. Magic angle spinning (MAS) DNP with electron decoupling requires a high electron Rabi frequency provided by a high-power microwave source such as a frequency-agile gyrotron. This dissertation describes the development …

Imaging The Three-Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri-Spot Point Spread Function, Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Fluorescence photons emitted by single molecules contain rich information regarding their rotational motions, but adapting single-molecule localization microscopy (SMLM) to measure their orientations and rotational mobilities with high precision remains a challenge. Inspired by dipole radiation patterns, we design and implement a Tri-spot point spread function (PSF) that simultaneously measures the three-dimensional orientation and the rotational mobility of dipole-like emitters across a large field of view. We show that the orientation measurements done using the Tri-spot PSF are sufficiently accurate to correct the anisotropy-based localization bias, from 30 nm to 7 nm, in SMLM. We further characterize the emission anisotropy …

Measuring 3d Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang, Tianben Ding, Jin Lu, Hesam Mazidi, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

We present a method to measure the molecular orientation and rotational mobility of single-molecule emitters by designing and implementing a Tri-spot point spread function. It can measure all degrees of freedom related to molecular orientation and rotational mobility. Its design is optimized by maximizing the theoretical limit of its measurement precision. We evaluate the precision and accuracy of the Tri-spot PSF by measuring the orientation and effective rotational mobility of single fluorescent molecules embedded in a polymer matrix.