京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第21732号 / 工博第4549号 / 新制||工||1709(附属図書館) / 京都大学大学院工学研究科電気工学専攻 / (主査)教授 大村 善治, 教授 松尾 哲司, 教授 竹内 繁樹 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

CHARLES UNIVERSITY and HASSELT UNIVERSITY / tUL Doctoral dissertation Collagen structures from cell culture to intact tendon ABSTRACT Author: Daniel Hadraba Promoters: Assoc. Prof. Karel Jelen | Charles University Prof. Marcel Ameloot | Hasselt University Co-promoters: Dr. Frantisek Lopot | Charles University Prof. Virginie Bito | Hasselt University Annotation Author: Ing. Mgr. Daniel Hadraba Doctoral thesis title: Collagen structures from cell culture to intact tendon Year: 2010 - 2017 Doctoral program: Doctor of Biomechanics at Charles University Doctor of Biomedical Science at Hasselt University / transnational University Limburg Departments: Dept. Anatomy and Biomechanics | Faculty of Physical Education and Sport | Charles University Dept. Biophysics | Hasselt University Promoters: Assoc. Prof. Karel Jelen | Dept. Anatomy and Biomechanics | Faculty of Physical Education and Sport | Charles University Prof. Marcel Ameloot | Hasselt University / transnational University Limburg Co-promoters: Dr. Frantisek Lopot | Dept. Anatomy and Biomechanics | Faculty of Physical Education and Sport | Charles University Prof. Virginie Bito | Hasselt University / transnational University Limburg Bibliography details: Pages 102 Figures 30 Tables 2 Equations 17 Keywords: tendon, collagen, crimps, orientation, aging,...

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Time Dependent Density Functional Theory offers a good accuracy/computational cost ratio among different methods used to predict the electronic structure for molecules of practical interest. The Coupled Electronic Oscillator (CEO) formalism was recently shown to accurately predict Nonlinear Optical (NLO) properties of organic chromophores when combined with Time Dependent Density Functional Theory. Unfortunately, CEO does not lend itself easily to interpretation of the structure activity relationships of chromophores. On the other hand, the Sum Over States formalism in combination with semiempirical wavefunction methods has been used in the past for the design of simplified essential states models. These models can be applied to optimization of NLO properties of interest for applications. Unfortunately, TD-DFT can not be combined directly with SOS because state-to-state transition dipoles are not defined in the linear response TD approach. In this work, a second order CEO approach to TD-DFT is simplified so that properties of double excited states and state-to-state transition dipoles may be expressed through the combination of linear response properties. This approach is termed the a posteriori Tamm-Dancoff approximation (ATDA), and validated against high-level wavefunction theory methods. Sum over States (SOS) and related Two-Photon Transition Matrix formalism are then used to predict Two-Photon Absorption (2PA) profiles and anisotropy, as well as Second Harmonic Generation (SHG) properties. Numerical results for several conjugated molecules are in excellent agreement with CEO and finite field calculations, and reproduce experimental measurements well.

No / This paper presents a circuit model which is used to calibrate the performance of nonlinear RF energy conversion inside a high quality factor resonant cavity with a known nonlinear loading device. The nonlinear radiofrequency energy conversion can be detected by exciting the fundamental operating frequency and observing the second harmonic resonant frequency within a doubly resonant cavity. By implementing the proposed mathematical model, the required input power can be estimated to maximise the chance of detecting the weak second harmonic signal prior to carry out the measurement.

Second harmonic (SH) microscopy represents a powerful tool for the investigation of crystalline systems, such as ferroelectrics and their domain walls (DWs). Under the condition of normal dispersion, i.e., the refractive index at the SH wavelength is larger as compared to the refractive index at the fundamental wavelength, n(2ω) . n(ω), bulk crystals will generate no SH signal. Should the bulk, however, contain DWs, an appreciable SH signal will still be detectable at the location of DWs stemming from the Čerenkov mechanism. In this work, we demonstrate both how SH signals are generated in bulk media and how the Čerenkov mechanism can be inhibited by using anomalous dispersion, i.e., n(ω) . n(2ω). This allows us to quantitatively estimate the relative strength of the Čerenkov compared to other SH contrast mechanisms in DWs, such as the interference contrast. The results are in agreement with previous experiments based on the geometric separation of the signals. Due to the observed, strong Čerenkov contrast, such signal contributions may not be neglected in polarimetry studies of ferroelectric DWs in the future.

The extracellular matrix (ECM) is a three-dimensional, acellular scaffold of living tissues. Incorporating the ECM into cell culture models is a goal of cell biology studies and requires biocompatible materials that can mimic the ECM. Among such materials are hydrogels: polymeric networks that derive most of their mass from water. With the tuning of their properties, these polymer networks can resemble living tissues. The microarchitectural properties of hydrogels, such as porosity, pore size, fiber length, and surface topology can determine cell plasticity. The adequate characterization of these parameters requires reliable and reproducible methods. However, most methods were historically standardized using other biological specimens, such as 2D cell cultures, biopsies, or even animal models. Therefore, their translation comes with technical limitations when applied to hydrogel-based cell culture systems. In our current work, we have reviewed the most common techniques employed in the characterization of hydrogel microarchitectures. Our review provides a concise description of the underlying principles of each method and summarizes the collective data obtained from cell-free and cell-loaded hydrogels. The advantages and limitations of each technique are discussed, and comparisons are made. The information presented in our current work will be of interest to researchers who employ hydrogels as platforms for cell culture, 3D bioprinting, and other fields within hydrogel-based research.

<p>The thesis consists of two main parts of nonlinear optical instrumentation development. </p> <p>Fluorescence-detected mid-infrared photothermal (F-PTIR) microscopy is demonstrated for sub-diffraction limited mid-infrared microspectroscopy of model systems and applied to probe phase transformations in amorphous solid dispersions. To overcome the diffraction limit in infrared imaging, a highly localized temperature-dependent photothermal effect is an attractive alternative indicator to infrared absorption. Photothermal atomic force microscopy infrared spectroscopy (AFM-IR) achieves nanometer resolution by monitoring heat caused expansion but only restricted on the surface. For 3D imaging, optically detected photothermal infrared (O-PTIR) combines an infrared laser with a visible probe source with to transduce photothermal refractive index changes (e.g., from changes in beam divergence or scattering). The sensitivity of O-PTIR is ultimately limited by the relatively weak dependence of refractive index with temperature, exhibiting changes of ~0.01% per oC. Fluorescence-detected photothermal mid-infrared (F-PTIR) spectroscopy (Fig. 1) is demonstrated herein to support 3D imaging with improved photothermal sensitivity. In F-FTIR, the sensitivity of fluorescence quantum efficiency to temperature change (~1-2% per oC) is used to transduce transient heat flux from localized IR absorption. The infrared spatial resolution of F-FTIR is defined by fluorescence microscopy and the thermal diffusivity of the sample instead of infrared wavelength. Initial F-PTIR proof of concept studies are described for microparticle assemblies of silica gel and polyethylene glycol, followed by applications of F-PTIR for analysis of localized composition within phase-separated domains induced by water vapor exposure of an amorphous solid dispersion (ritonavir in copovidone).</p> <p>Fluorescence recovery while photobleaching (FRWP) is demonstrated as a method for quantitative measurements of rapid diffusion mapping over the microsecond to millisecond time scale. Diffusion measurements are critical for molecular mobility assessment in cell biology, materials science and pharmacology. Fluorescence recovery after photobleaching (FRAP) is a well-known noninvasive optical microscopy method for measuring diffusion coefficients of macromolecules, such as proteins in cells and viscous solutions. However, conventional point-bleach FRAP is challenging to implement with multi-photon excitation and typically only supports diffusion analysis over millisecond time scales due to camera frame rate limitations. FRWP with patterned illumination addresses these limitations of FRAP by probing the fluorescence intensity changes while bleaching a comb pattern within a field of view (FoV). Fast-scanning of an ultrafast excitation beam distributes heat rapidly over multiple adjacent pixels, minimizing local heating effects that could complicate analogous diffusion measurements by point-bleach FRAP with multiphoton excitation. In FRWP, time-scales of the probed diffusion events are defined by a single line-pass time of a resonant scanning-mirror with a period of 125  s. In FRWP, the bleach pattern spans locations across the whole FoV, enabling diffusion mapping through image segmentation. More than a hundred bleaching and recovery events can be recorded during a single 10s measurement. Normal and anomalous diffusion of rhodamine-labeled bovine serum albumin (BSA) molecules was studied as a model system, with applications targeting rapid assessment of therapeutic macromolecule mobility within heterogeneous biological environments.</p>

<p>Space-based global navigation and precision timing systems are critical for modern infrastructure. Atomic clock technology has increased the precision of these systems so that they are viable for military operations, navigation, telecommunications, and finance. Advances in optical atomic clocks, based on optical frequencies, provide an opportunity for even more precise timing. Therefore, developments in chip-scale optical atomic clock technologies could lead to increased and more wide-spread application of this precision timing. One component of the optical atomic clock is the optical frequency comb which serves as an interface between optical and microwave frequencies. This thesis will cover experiments related to these optical frequency combs. A 2$\mu$m fiber laser was developed in order to test second harmonic devices required to stabilize an optical frequency comb. The laser was then employed to measure the operating wavelengths and efficiencies of non-linear devices. In addition, an analysis of the radio frequency instruments used to evaluate microwave outputs was conducted to determine whether a digital signal analyzer (oscilloscope) or an analog electronic spectrum analyzer provides more accurate results for optical frequency comb based experiments.</p>