From molecules to organs : Microscopy and multi-scale nature of development

Morphogenesis is a phenomenon of intricate balance and dynamic interplay between processes occurring at a wide range of scales (spatial, temporal and energetic). During development, a variety of physical mechanisms are employed by tissues to simultaneously pattern, move, and differentiate based on information exchange between constituent cells, perhaps more than at any other time during an organism's life. To fully understand such events, a combined theoretical and experimental framework is required to assist in deciphering the correlations at both structural and functional levels at scales that include the intracellular and tissue levels as well as organs and organ systems. Microscopy, especially diffraction-limited light microscopy, has emerged as a central tool to capture the spatio-temporal context of life processes. Imaging has the unique advantage of watching biological events as they unfold over time at single-cell resolution in the intact animal. In this work I present a range of problems in morphogenesis, each unique in its requirements for novel quantitative imaging both in terms of the technique and analysis. Understanding the molecular basis for a developmental process involves investigating how genes and their products- mRNA and proteins-function in the context of a cell. Structural information holds the key to insights into mechanisms and imaging fixed specimens paves the first step towards deciphering gene function. The work presented in this thesis starts with the demonstration that the fluorescent signal from the challenging environment of whole-mount imaging, obtained by in situ hybridization chain reaction (HCR), scales linearly with the number of copies of target mRNA to provide quantitative sub-cellular mapping of mRNA expression within intact vertebrate embryos. The work then progresses to address aspects of imaging live embryonic development in a number of species. While processes such as avian cartilage growth require high spatial resolution and lower time resolution, dynamic events during zebrafish somitogenesis require higher time resolution to capture the protein localization as the somites mature. The requirements on imaging are even more stringent in case of the embryonic zebrafish heart that beats with a frequency of ~ 2-2.5 Hz, thereby requiring very fast imaging techniques based on two-photon light sheet microscope to capture its dynamics. In each of the hitherto-mentioned cases, ranging from the level of molecules to organs, an imaging framework is developed, both in terms of technique and analysis to allow quantitative assessment of the process in vivo. Overall the work presented in this thesis combines new quantitative tools with novel microscopy for the precise understanding of processes in embryonic development.

Optical intrinsically fuzzy mathematical morphology for gray-scale image processing

Intrinsically fuzzy morphological erosion and dilation are extended to a total of eight operations that have been formulated in terms of a single morphological operation--biased dilation. Based on the spatial coding of a fuzzy variable, a bidirectional projection concept is proposed. Thus, fuzzy logic operations, arithmetic operations, gray-scale dilation, and erosion for the extended intrinsically fuzzy morphological operations can be included in a unified algorithm with only biased dilation and fuzzy logic operations. To execute this image algebra approach we present a cellular two-layer processing architecture that consists of a biased dilation processor and a fuzzy logic processor. (C) 1996 Optical Society of America

Morphological ordered gray-scale erosion for optical image processing

An ordered gray-scale erosion is suggested according to the definition of hit-miss transform. Instead of using three operations, two images, and two structuring elements, the developed operation requires only one operation and one structuring element, but with three gray-scale levels. Therefore, a union of the ordered gray-scale erosions with different structuring elements can constitute a simple image algebra to program any combined image processing function. An optical parallel ordered gray-scale erosion processor is developed based on the incoherent correlation in a single channel. Experimental results are also given for an edge detection and a pattern recognition. (C) 1998 Society of Photo-Optical Instrumentation Engineers. [S0091-3286(98)00306-7].

Fabrication, Characterization, And Deformation of 3D Structural Meta-Materials

Current technological advances in fabrication methods have provided pathways to creating architected structural meta-materials similar to those found in natural organisms that are structurally robust and lightweight, such as diatoms. Structural meta-materials are materials with mechanical properties that are determined by material properties at various length scales, which range from the material microstructure (nm) to the macro-scale architecture (μm – mm). It is now possible to exploit material size effect, which emerge at the nanometer length scale, as well as structural effects to tune the material properties and failure mechanisms of small-scale cellular solids, such as nanolattices. This work demonstrates the fabrication and mechanical properties of 3-dimensional hollow nanolattices in both tension and compression. Hollow gold nanolattices loaded in uniaxial compression demonstrate that strength and stiffness vary as a function of geometry and tube wall thickness. Structural effects were explored by increasing the unit cell angle from 30° to 60° while keeping all other parameters constant; material size effects were probed by varying the tube wall thickness, t, from 200nm to 635nm, at a constant relative density and grain size. In-situ uniaxial compression experiments reveal an order-of-magnitude increase in yield stress and modulus in nanolattices with greater lattice angles, and a 150% increase in the yield strength without a concomitant change in modulus in thicker-walled nanolattices for fixed lattice angles. These results imply that independent control of structural and material size effects enables tunability of mechanical properties of 3-dimensional architected meta-materials and highlight the importance of material, geometric, and microstructural effects in small-scale mechanics. This work also explores the flaw tolerance of 3D hollow-tube alumina kagome nanolattices with and without pre-fabricated notches, both in experiment and simulation. Experiments demonstrate that the hollow kagome nanolattices in uniaxial tension always fail at the same load when the ratio of notch length (a) to sample width (w) is no greater than 1/3, with no correlation between failure occurring at or away from the notch. For notches with (a/w) > 1/3, the samples fail at lower peak loads and this is attributed to the increased compliance as fewer unit cells span the un-notched region. Finite element simulations of the kagome tension samples show that the failure is governed by tensile loading for (a/w) < 1/3 but as (a/w) increases, bending begins to play a significant role in the failure. This work explores the flaw sensitivity of hollow alumina kagome nanolattices in tension, using experiments and simulations, and demonstrates that the discrete-continuum duality of architected structural meta-materials gives rise to their flaw insensitivity even when made entirely of intrinsically brittle materials.

Optical multichannel-scale range-tunable Fourier-transforming system: experiment

The authors have demonstrated the principle of a novel optical multichannel-scale range-tunable Fourier-transforming system. The experimental results show good agreement with the theoretical analysis.

Aerodynamics of vertical-axis wind turbines in full-scale and laboratory-scale experiments

Within a wind farm, multiple turbine wakes can interact and have a substantial effect on the overall power production. This makes an understanding of the wake recovery process critically important to optimizing wind farm efficiency. Vertical-axis wind turbines (VAWTs) exhibit features that are amenable to dramatically improving this efficiency. However, the physics of the flow around VAWTs is not well understood, especially as it pertains to wake interactions, and it is the goal of this thesis to partially fill this void. This objective is approached from two broadly different perspectives: a low-order view of wind farm aerodynamics, and a detailed experimental analysis of the VAWT wake.

One of the contributions of this thesis is the development of a semi-empirical model of wind farm aerodynamics, known as the LRB model, that is able to predict turbine array configurations to leading order accuracy. Another contribution is the characterization of the VAWT wake as a function of turbine solidity. It was found that three distinct regions of flow exist in the VAWT wake: (1) the near wake, where periodic blade shedding of vorticity dominates; (2) a transition region, where growth of a shear-layer instability occurs; (3) the far wake, where bluff-body oscillations dominate. The wake transition can be predicted using a new parameter, the dynamic solidity, which establishes a quantitative connection between the wake of a VAWT and that of a circular cylinder. The results provide insight into the mechanism of the VAWT wake recovery and the potential means to control it.

Measuring temporal fluctuation on femtosecond scale in a random medium

We report an observation of femtosecond optical fluctuations of transmitted light when a coherent femtosecond pulse propagates through a random medium. They are a result of random interference among scattered waves coming from different trajectories in the time domain. Temporal fluctuations are measured by using cross-correlated frequency optical gating. It is shown that a femtosecond pulse will be broadened and distorted in pulse shape while it is propagating in random medium. The real and imaginary components of transmitted electric field are also distorted severely. The average of the fluctuated transmission pulses yields a smooth profile, probability functions show good agreement with Gaussian distribution. (c) 2007 Elsevier B.V. All rights reserved.

Non-Linear Scale Interactions in a Forced Turbulent Boundary Layer

This thesis explores the dynamics of scale interactions in a turbulent boundary layer through a forcing-response type experimental study. An emphasis is placed on the analysis of triadic wavenumber interactions since the governing Navier-Stokes equations for the flow necessitate a direct coupling between triadically consist scales. Two sets of experiments were performed in which deterministic disturbances were introduced into the flow using a spatially-impulsive dynamic wall perturbation. Hotwire anemometry was employed to measure the downstream turbulent velocity and study the flow response to the external forcing. In the first set of experiments, which were based on a recent investigation of dynamic forcing effects in a turbulent boundary layer, a 2D (spanwise constant) spatio-temporal normal mode was excited in the flow; the streamwise length and time scales of the synthetic mode roughly correspond to the very-large-scale-motions (VLSM) found naturally in canonical flows. Correlation studies between the large- and small-scale velocity signals reveal an alteration of the natural phase relations between scales by the synthetic mode. In particular, a strong phase-locking or organizing effect is seen on directly coupled small-scales through triadic interactions. Having characterized the bulk influence of a single energetic mode on the flow dynamics, a second set of experiments aimed at isolating specific triadic interactions was performed. Two distinct 2D large-scale normal modes were excited in the flow, and the response at the corresponding sum and difference wavenumbers was isolated from the turbulent signals. Results from this experiment serve as an unique demonstration of direct non-linear interactions in a fully turbulent wall-bounded flow, and allow for examination of phase relationships involving specific interacting scales. A direct connection is also made to the Navier-Stokes resolvent operator framework developed in recent literature. Results and analysis from the present work offer insights into the dynamical structure of wall turbulence, and have interesting implications for design of practical turbulence manipulation or control strategies.

Small scale turbulence in high Karlovitz number premixed flames

The purpose of this thesis is to characterize the behavior of the smallest turbulent scales in high Karlovitz number (Ka) premixed flames. These scales are particularly important in the two-way coupling between turbulence and chemistry and better understanding of these scales will support future modeling efforts using large eddy simulations (LES). The smallest turbulent scales are studied by considering the vorticity vector, ω, and its transport equation.

Due to the complexity of turbulent combustion introduced by the wide range of length and time scales, the two-dimensional vortex-flame interaction is first studied as a simplified test case. Numerical and analytical techniques are used to discern the dominate transport terms and their effects on vorticity based on the initial size and strength of the vortex. This description of the effects of the flame on a vortex provides a foundation for investigating vorticity in turbulent combustion.

Subsequently, enstrophy, ω² = ω • ω, and its transport equation are investigated in premixed turbulent combustion. For this purpose, a series of direct numerical simulations (DNS) of premixed n-heptane/air flames are performed, the conditions of which span a wide range of unburnt Karlovitz numbers and turbulent Reynolds numbers. Theoretical scaling analysis along with the DNS results support that, at high Karlovitz number, enstrophy transport is controlled by the viscous dissipation and vortex stretching/production terms. As a result, vorticity scales throughout the flame with the inverse of the Kolmogorov time scale, τ_η, just as in homogeneous isotropic turbulence. As τ_η is only a function of the viscosity and dissipation rate, this supports the validity of Kolmogorov’s first similarity hypothesis for sufficiently high Ka numbers (Ka ≳ 100). These conclusions are in contrast to low Karlovitz number behavior, where dilatation and baroclinic torque have a significant impact on vorticity within the flame. Results are unaffected by the transport model, chemical model, turbulent Reynolds number, and lastly the physical configuration.

Next, the isotropy of vorticity is assessed. It is found that given a sufficiently large value of the Karlovitz number (Ka ≳ 100) the vorticity is isotropic. At lower Karlovitz numbers, anisotropy develops due to the effects of the flame on the vortex stretching/production term. In this case, the local dynamics of vorticity in the strain-rate tensor, S, eigenframe are altered by the flame. At sufficiently high Karlovitz numbers, the dynamics of vorticity in this eigenframe resemble that of homogeneous isotropic turbulence.

Combined, the results of this thesis support that both the magnitude and orientation of vorticity resemble the behavior of homogeneous isotropic turbulence, given a sufficiently high Karlovitz number (Ka ≳ 100). This supports the validity of Kolmogorov’s first similarity hypothesis and the hypothesis of local isotropy under these condition. However, dramatically different behavior is found at lower Karlovitz numbers. These conclusions provides/suggests directions for modeling high Karlovitz number premixed flames using LES. With more accurate models, the design of aircraft combustors and other combustion based devices may better mitigate the detrimental effects of combustion, from reducing CO₂ and soot production to increasing engine efficiency.