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Payton, Oliver David. "High-speed atomic force microscopy under the microscope." Thesis, University of Bristol, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.574416.
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SINCE its invention in 1986, the atomic force microscope (AFM) has revolutionised the field of nanotechnology and nanoscience. It is a tool that has enabled research into areas of medicine, advanced materials, biology, chemistry and physics. However due to its low frame rate it is a tool that has been limited to imaging small areas using a time lapse technique. It has only been in recent years that the frame rate of the device has been increased in a tool known as high-speed AFM (HSAFM). This increased frame rate allows, for the first time, biological processes to be viewed in real time or macro sized areas to be imaged with nanoscale resolution. The research presented here concentrates on a specific type of high-speed AFM developed at the University of Bristol called contact mode HSAFM. This thesis explains how the microscope is able to function, and presents a leap in image quality due to an increased understanding of the dynamics of the system. The future of the device is also discussed. III
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Franklin, Thomas. "Scanning ionoluminescence microscopy with a helium ion microscope." Thesis, University of Southampton, 2012. https://eprints.soton.ac.uk/352281/.
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The ORIONR PLUS scanning helium ion microscope (HIM) images at sub nanometer resolution. Images of the secondary electron emission have superior resolution and depth of field compared to a scanning electron microscope (SEM). Ionoluminescent imaging is not an area that has been extensively explored by typical ion beam systems as they have large spot sizes in the region of microns, leading to poor spatial resolution. This thesis confirms that the ORIONR PLUS can form images from the ionoluminescent signal, resolutions of 20nm can be obtained for images of bright nanoparticles. Ionoluminescence spectra can also be obtained from some samples. The position of emission peaks in samples under the ORIONR PLUS does not deviate significantly from cathodoluminescence (CL) peaks under SEM. However, the relative heights of the emission peaks in a sample can vary between ionoluminescence (IL) and CL. In addition, It is found that there exists a proportional relationship between acceleration voltage and ionoluminescent signal in the ORIONR PLUS, this relationship is also exhibited in CL. However, when normalised for current and acceleration voltage there appears to be no samples that show greater luminescence under ionoluminescence than cathodoluminescence, with ionoluminescent intensities up to an order of magnitude lower. Ionoluminescence under the ORIONR PLUS is found to be a poor candidate for the analysis of direct band gap semiconductors, this is attributed to the smaller interaction volumes and achievable beam current of the ORIONR PLUS. It is also found that some direct band gap materials are very susceptible to beam damage under the ion beam at beam doses typically used for secondary electron (SE) imaging. It is possible to obtain simultaneous IL and SE images of organic fluorospores in a biological sample. However, the luminescence of the fluorospores was only just sufficient to form images with a 200nm resolution. Rare earth based nanoparticles show brighter luminescence and greater resistance to beam damage than organic fluorospores. If such particles could be utilised for immunofluorescence it would make combined secondary electron and immunofluorescence imaging under the ORIONR PLUS a viable technique.
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Szelc, Jedrzej. "THz imaging and microscopy : a multiplexed near-field TeraHertz microscope." Thesis, University of Southampton, 2011. https://eprints.soton.ac.uk/209643/.
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Wright, Adele Hart. "Design, development, and application of an automated precision scanning microscope stage with a controlled environment." Thesis, Georgia Institute of Technology, 1997. http://hdl.handle.net/1853/16409.
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Battistella, Eliana. "Towards an improved photonic force microscope: a novel technique for biological microscopy." Master's thesis, Alma Mater Studiorum - Università di Bologna, 2017. http://amslaurea.unibo.it/14864/.
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Una delle tecniche più note nello studio topografico di campioni biologici è l’AFM. Ci sono però limitazioni dovute alla presenza del cantilever, il quale pone un limite nella forza minima applicabile su un campione per ottenere un’immagine topografica. Questa forza (ordine dei 10 pN) può essere sufficiente a danneggiare il campione e a deformare i dettagli topografici che si vorrebbero evidenziare. Per superare questo problema si può usare un Photonic Force Microscope, dove il cantilever è sostituito da Optical Tweezers. Questa tecnica permette di effettuare scansioni di campioni biologici applicando forze dell’ordine dei 100 fN. All’interno della trappola ottica viene posizionata una microparticella che agisce da sonda, attraverso la quale possono essere rilevati dettagli topografici del campione. La differenza rispetto al PFM tradizionale si trova proprio nel tipo di sonda utilizzata durante la scansione. Lo standard prevede l’utilizzo di una sonda sferica, di dimensioni dell’ordine dei 100 nm mentre l’ipotesi è che si possano utilizzare delle sonde cilindriche con alla base un dettaglio acuminato che richiama la punta dell’AFM. Questo tipo di sonda consentirebbe di raggiungere una risoluzione maggiore, rispetto al PFM tradizionale, che risente del limite dato dal diametro della sfera. Due differenti setup per la PFM sono stati costruiti e testati durante questo periodo di tesi. Sono state testate diverse microparticelle cilindriche, di dimensioni differenti in termini di aspect ratio con lo scopo di osservare la stabilità di questo tipo di sonda. Nei risultati viene proposto un metodo per controllare la stabilità e l’orientazione della microparticella cilindrica all’interno della trappola ottica. Viene inoltre fatta un’ipotesi su un metodo per stimare la lunghezza della punta che dovrà essere verificata da una misura sistematica. I risultati preliminari riguardanti la scansione di strutture note suggeriscono la validità dell’uso di questo nuovo tipo di sonda.
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Bethge, Philipp. "Development of a two-photon excitation STED microscope and its application to neuroscience." Thesis, Bordeaux, 2014. http://www.theses.fr/2014BORD0018/document.
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L’avènement de la microscopie STED (Stimulated Emission Depletion) a bouleversé le domaine desneurosciences du au fait que beaucoup de structures neuronale, tels que les épines dendritiques, lesaxones ou les processus astrocytaires, ne peuvent pas être correctement résolu en microscopiephotonique classique. La microscopie 2-photon est une technique d’imagerie photonique très largement utilisée dans le domaine des neurosciences car elle permet d’imager les événements dynamique en profondeur dans le tissu cérébral, offrant un excellent sectionnement optique et une meilleure profondeur de pénétration. Cependant, la résolution spatiale de cette approche est limitée autour de 0.5 μm, la rendant inappropriée pour étudier les détails morphologiques des neurones et synapses. Le but de mon travail de thèse était à A) développer un microscope qui permet d'améliorer l'imagerie 2-photon en la combinant avec la microscopie STED et B) démontrer son potentiel pour l'imagerie à l'échelle nanométrique de processus neuronaux dynamiques dans des tranches de cerveau aigus et in vivo. Le nouveau microscope permet d'obtenir une résolution spatiale latérale de ~ 50 nm à des profondeurs d'imagerie de ~ 50 μm dans du tissu cérébral vivant. Il fonctionne avec des fluorophores verts, y compris les protéines fluorescentes communes telles que la GFP et YFP, offrant le contraste de deux couleurs basé sur la détection spectrale et linéaire ‘unmixing’. S’agissant d’un microscope droit, utilisant un objectif à immersion ayant une grande distance de travail, nous avons pu incorporer des techniques électrophysiologiques comme patch-clamp et ajouter une plateforme pour l'imagerie in vivo. J’ai utilise ce nouveau microscope pour imager des processus neuronaux fins et leur dynamique à l’échelle nanométrique dans différent types de préparations et des régions différentes du cerveau. J’ai pu révéler des nouvelles caractéristiques morphologique des dendrites et épines. En outre, j'ai exploré différentes stratégies de marquage pour pouvoir utiliser la microscopie STED pour imager le trafic des protéines et de leur dynamique à l'échelle nanométrique dans des tranches de cerveau<br>The advent of STED microscopy has created a lot of excitement in the field of neuroscience becausemany important neuronal structures, such as dendritic spines, axonal shafts or astroglial processes,cannot be properly resolved by regular light microscopy techniques. Two-photon fluorescence microscopy is a widely used imaging technique in neuroscience because it permits imaging dynamic events deep inside light-scattering brain tissue, providing high optical sectioning and depth penetration. However, the spatial resolution of this approach is limited to around half a micron, and hence is inadequate for revealing many morphological details of neurons and synapses. The aim of my PhD work was to A) develop a microscope that improves on two-photon imaging by combining it with STED microscopy and to B) demonstrate its potential for nanoscale imaging of dynamic neural processes in acute brain slices and in vivo. The new microscope achieves a lateral spatial resolution of ~50 nm at imaging depths of ~50 μm in living brain slices. It works with green fluorophores, including common fluorescent proteins like GFP and YFP, offering two-color contrast based on spectral detection and linear unmixing. Because of its upright design using a long working distance water-immersion objective, it was possible to incorporate electrophysiological techniques like patch-clamping or to add a stage for in vivo imaging. I have used the new microscope to image fine neural processes and their nanoscale dynamics in different experimental preparations and brain regions, revealing new and interesting morphological features of dendrites and spines. In addition, I have explored different labeling strategies to be able to use STED microscopy for visualizing protein trafficking and dynamics at the nanoscale in brain slices
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Yu, Enhua. "Crossed and uncrossed retinal fibres in normal and monocular hamsters : light and electron microscopic studies /." [Hong Kong : University of Hong Kong], 1990. http://sunzi.lib.hku.hk/hkuto/record.jsp?B13014316.
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Konda, Pavan Chandra. "Multi-Aperture Fourier Ptychographic Microscopy : development of a high-speed gigapixel coherent computational microscope." Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/9015/.
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Medical research and clinical diagnostics require imaging of large sample areas with sub-cellular resolution. Conventional imaging techniques can provide either high-resolution or wide field-of-view (FoV) but not both. This compromise is conventionally defeated by using a high NA objective with a small FoV and then mechanically scan the sample in order to acquire separate images of its different regions. By stitching these images together, a larger effective FoV is then obtained. This procedure, however, requires precise and expensive scanning stages and prolongs the acquisition time, thus rendering the observation of fast processes/phenomena impossible. A novel imaging configuration termed Multi-Aperture Fourier Ptychographic Microscopy (MA-FPM) is proposed here based on Fourier ptychography (FP), a technique to achieve wide-FoV and high-resolution using time-sequential synthesis of a high-NA coherent illumination. MA-FPM configuration utilises an array of objective lenses coupled with detectors to increase the bandwidth of the object spatial-frequencies captured in a single snapshot. This provides high-speed data-acquisition with wide FoV, high-resolution, long working distance and extended depth-of-field. In this work, a new reconstruction method based on Fresnel diffraction forward model was developed to extend FP reconstruction to the proposed MA-FPM technique. MA-FPM was validated experimentally by synthesis of a 3x3 lens array system from a translating objective-detector system. Additionally, a calibration procedure was also developed to register dissimilar images from multiple cameras and successfully implemented on the experimental data. A nine-fold improvement in captured data-bandwidth was demonstrated. Another experimental configuration was proposed using the Scheimpflug condition to correct for the aberrations present in the off-axis imaging systems. An experimental setup was built for this new configuration using 3D printed parts to minimise the cost. The design of this setup is discussed along with robustness analysis of the low-cost detectors used in this setup. A reconstruction model for the Scheimpflug configuration FP was developed and applied to the experimental data. Preliminary experimental results were found to be in agreement with this reconstruction model. Some artefacts were observed in these results due to the calibration errors in the experiment. These can be corrected by using the self-calibration algorithm proposed in the literature, which is left as a future work. Extensions to this work can include implementing multiplexed illumination for further increasing the data acquisition speed and diffraction tomography for imaging thick samples.
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Morgan, Scott Warwick. "Gaseous secondary electron detection and cascade amplification in the environmental scanning electron microscope /." Electronic version, 2005. http://adt.lib.uts.edu.au/public/adt-NTSM20060511.115302/index.html.
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Morrish, Dru, and DruMorrish@gmail com. "Morphology dependent resonance of a microscope and its application in near-field scanning optical microscopy." Swinburne University of Technology. Centre for Micro-Photonics, 2005. http://adt.lib.swin.edu.au./public/adt-VSWT20051124.121838.
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In recent times, near-field optical microscopy has received increasing attention for its
ability to obtain high-resolution images beyond the diffraction limit. Near-field optical
microscopy is achieved via the positioning and manipulation of a probe on a scale less
than the wavelength of the incident light.
Despite many variations in the mechanical design of near-field optical microscopes almost all rely on direct mechanical access of a cantilever or a derivative form to probe the sample. This constricts the study to surface examinations in simple sample environments. Distance regulation between the sample surface and the delicate probe
requires its own feedback mechanism. Determination of feedback is achieved through
monitoring the shift of resonance of one arm of a 'tuning fork', which is caused by the interaction of the probes tip with the Van der Waals force. Van der Waals force emanates from atom-atom interaction at the top of the sample surface. Environmental contamination of the sample surface with additional molecules such as water makes accurate measurement of these forces particularly challenging. The near-field study of
living biological material is extremely difficult as an aqueous environment is required for its extended survival. Probe-sample interactions within an aqueous environment that
result in strong detectable signal is a challenging problem that receives considerable
attention and is a focus of this thesis.
In order to increase the detectible signal a localised field enhancement in the probing region is required. The excitation of an optically resonant probe by morphology dependent resonance (MDR) provides a strong localised field enhancement. Efficient MDR excitation requires important coupling conditions be met, of which the localisation of the incident excitation is a critical factor.
Evanescent coupling by frustrated total internal reflection to a MDR microcavity provides an ideal method for localised excitation. However it has severe drawbacks if the probe is to be manipulated in a scanning process. Tightly focusing the incident illumination by a high numerical aperture objective lens provides the degree of freedom to enable both MDR excitation and remote manipulation. Two-photon nonlinear excitation is shown to couple efficiently to MDR modes due to the high spatial localisation of the incident excitation in three-dimensions. The dependence of incident excitation localisation by high numerical aperture objective on MDR efficiency is thoroughly examined in this thesis. The excitation of MDR can be enhanced by up to 10 times with the localisation of the incident illumination from the centre of the
microcavity to its perimeter.
Illuminating through a high numerical aperture objective enables the remote noninvasive
manipulation of a microcavity probe by laser trapping. The transfer of photon momentum from the reflection and refraction of the trapping beam is sufficient enough to exert piconewtons of force on a trapped particle. This allows the particle to be held and scanned in a predictable fashion in all three-dimensions. Optical trapping
removes the need for invasive mechanical access to the sample surface and provides a means of remote distance regulation between the trapped probe and the sample. The femtosecond pulsed beam utilised in this thesis allows the simultaneous induction of two-photon excitation and laser trapping. It is found in this thesis that a MDR microcavity can be excited and translated in an efficient manner. The application of this technique to laser trapped near-field microscopy and single molecule detection is of particular interest.
Monitoring the response of the MDR signal as it is scanned over a sample object enables a near-field image to be built up. As the enhanced evanescent field from the propagation of MDR modes around a microcavity interacts with different parts of the sample, a measurable difference in energy leakage from the cavity modes occurs. The definitive spectral properties of MDR enables a multidimensional approach to imaging and sensing, a focus of this thesis. Examining the spectral modality of the MDR signal
can lead to a contrast enhancement in laser trapped imaging. Observing a single MDR mode during the scanning process can increase the image contrast by up to 1:23 times compared to that of the integrated MDR fluorescence spectrum.
The work presented in this thesis leads to the possibility of two-photon fluorescence
excitation of MDR in combination with laser trapping becoming a valuable tool in near-
field imaging, sensing and single molecule detection in vivo. It has been demonstrated
that particle scanned, two-photon fluorescence excitation of MDR, by laser trapping 'tweezers' can provide a contrast enhancement and multiple imaging modalities. The spectral imaging modality has particular benefits for image contrast enhancements.
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PONZIO, FRANCESCO. "Deep learning at the microscope - Working towards improved microscopy image analysis with deep neural networks." Doctoral thesis, Politecnico di Torino, 2021. http://hdl.handle.net/11583/2935596.
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Eubanks, Craig S. "Comparison of ellipso-polarimetry and dark-field methods for determination of thickness variations in thin films /." Online version of thesis, 1992. http://hdl.handle.net/1850/11269.
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于恩華 and Enhua Yu. "Crossed and uncrossed retinal fibres in normal and monocular hamsters: light and electron microscopic studies." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1990. http://hub.hku.hk/bib/B31232449.
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El, Hajraoui Khalil. "Études in-situ dans un microscope électronique en transmission des réactions à l’état solide entre métal et nanofil de Ge." Thesis, Université Grenoble Alpes (ComUE), 2017. http://www.theses.fr/2017GREAY012/document.
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Le domaine des nanofils semi-conducteurs est en pleine expansion depuis ces dix dernières années grâce à leurs applications dans de nombreux domaines tels que l’électronique ou la conversion d’énergie. Dans cette étude on part d’une base de nanofil de germanium (le canal), on dépose des contacts métalliques qui seront chauffés par effet joule. Une différence de potentiel est alors appliquée au contact d’entrée (la source), le courant électrique est récupéré et mesuré par le contact de sortie (le drain). Une réaction à l’état solide permet aux atomes du métal de diffuser dans le nanofil. La propagation d'une phase métal/semi-conducteur est suivie dans un microscope électronique en transmission (MET) dont la résolution permet une observation à l’échelle atomique au niveau de la source, le drain et le canal. Les dispositifs caractérisés au cours de ce stage ont été élaborés à partir de deux types de membranes, l’une plane et l’autre avec des trous. Chacune d’entre elles sont constituées d’une couche de nitrate de silicium Si3N4 à leurs surfaces présentant l’avantage d’être transparents aux électrons et isolants au courant<br>Semiconductor nanowires (NWs) are promising candidates for many device applications ranging from electronics and optoelectronics to energy conversion and spintronics. However, typical NW devices are fabricated using electron beam lithography and therefore source, drain and channel length still depend on the spatial resolution of the lithography. In this work we show fabrication of NW devices in a transmission electron microscope (TEM) where we can obtain atomic resolution on the channel length using in-situ propagation of a metallic phase in the semiconducting NW independent of the lithography resolution. We show results on semiconducting NW devices fabricated on two different electron transparent Si3N4 membranes: a planar membrane and a membrane where devices are suspended over holes. First we show the process of making lithographically defined reliable electrical contacts on individual NWs. Second we show first results on in-situ propagation of a metal-semiconductor phase in Ge NWs by joule heating, while measuring the current through the device. Two different devices are studied: one with platinum metal contacts and one with copper contacts. Different phenomena can occur in CuGe NWs during phase propagation
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Travaillot, Thomas. "Caractérisation mécanique des matériaux élastiques à l'échelle locale par microscopie à pointe vibrante : Approche multimodale et mesure de champs." Thesis, Besançon, 2014. http://www.theses.fr/2014BESA2011/document.
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Ces travaux de thèse proposent une amélioration du Scanning Microdeformation Microscope (SMM),un microscope à sonde locale, pour la caractérisation mécanique élastique des matériaux à l’échellelocale. Il est montré qu’en utilisant n > 2 modes de résonance du SMM, il est possible de découplerles mesures du module de Young et du coefficient de Poisson d’un matériau isotrope.Une étude des modes du résonateur a permis d’enrichir son modèle afin qu’il puisse modélisern > 2 modes. Des procédures ont été développées pour identifier les paramètres de ce modèle etles constantes élastiques des matériaux à partir de n > 2 fréquences de résonance. Enfin, ces procéduresont été appliquées à des exemples de caractérisation à l’échelle locale afin de valider laméthode et d’en exhiber les possibilités et les limites.Pour gagner en robustesse et se diriger vers la caractérisation des matériaux anisotropes, un systèmed’imagerie interférométrique en lumière polarisée, permettant la mesure du champ de rotationde surfaces réfléchissantes dans une direction particulière, a été développé pour être intégré auSMM. Son prisme biréfringent à gradient uniaxial d’indice lui confère sa sensibilité à la rotation. Cesystème est capable de mesurer un champ de rotation localisé comme c’est le cas au voisinage dela pointe du SMM. Il a aussi montré son intérêt dans les cas où l’effet d’échelle rend particulièrementintéressante la mesure de la rotation<br>This work proposes an improvement of the Scanning Microdeformation Microscope (SMM), a scanningprobe microscope, for the mechanical elastic characterization of materials at local scale. It demonstratesthat using n > 2 SMM resonance frequencies allows to decouple Young’s modulus andPoisson’s ratio values for an isotropic material.The mechanical description of the resonator has been enriched in order to allow for an accuratemodeling over a wide frequency range. Procedures have been developed to identify the modellingparameters and the elastic constants of the materials from n > 2 resonant frequencies. Finally, theseprocedures have been applied to the characterization of various materials at local scale in order tovalidate the method and to present possibilities and limits.To improve robustness and move towards the characterization of anisotropic materials, a polarizedlightimaging interferometer was developed to measure the rotation field of reflecting surfaces in aparticular direction. The sensitivity to the rotation originates from a homemade birefringent prism withuniaxial gradient of refractive index. This system is able to measure a localized rotation field as it isinduced in the vicinity of the tip of the SMM. Its interest is also demonstrated in cases in which scaleeffects make the rotation measurement preferable to the out-of-plane displacement measurement
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Bélisle, Jonathan. "Design and assembly of a multimodal nonlinear laser scanning microscope." Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=100765.
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The objective of this thesis is to present the fabrication of a multiphoton microscope and the underlying theory responsible for its proper functioning. A basic introduction to nonlinear optics will give the necessary knowledge to the reader to understand the optical effects involved. Femtosecond laser pulses will be presented and characterized. Each part of the microscope, their integration and the design of the microscope will be discussed. The basic concepts of laser scanning microscopy are also required to explain the design of the scanning optics. Fast scanning problems and their solutions are also briefly viewed. As a working proof, the first images taken with the microscope will be presented. Fluorescent beads, rat tail tendon, gold nanoparticles and pollen grain images using various nonlinear effects will be shown and discussed.
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Romero, Leiro Freddy José. "Poly-articulated microrobotics for correlative AFM-in-SEM microscopy." Electronic Thesis or Diss., Sorbonne université, 2023. https://accesdistant.sorbonne-universite.fr/login?url=https://theses-intra.sorbonne-universite.fr/2023SORUS520.pdf.
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La microscopie corrélative est le résultat de la combinaison de deux ou plusieurs techniques de microscopie pour fournir des informations complémentaires sur un échantillon. En utilisant un microscope électronique à balayage (MEB) et un microscope à force atomique (AFM), la microscopie corrélative AFM-in-SEM permet non seulement la caractérisation 3D d'échantillons observés à l'intérieur d'un MEB, mais aussi la manipulation de micro- et nanostructures avec une très grande précision. Cette technique peut être appliquée à divers échantillons dans les domaines de la biologie, de l'électronique et de la science des matériaux. Bien que les solutions AFM-in-SEM existantes dans l'état actuel soient puissantes, elles nécessitent des utilisateurs experts, elles ne sont pas assez polyvalentes pour être utilisées pour différents types de tâches et elles utilisent des robots AFM cartésiens qui limitent fortement la dextérité et la performance du système d'imagerie. L'objectif de cette thèse est d'étudier et d'expérimenter un concept original d'AFM basé sur la robotique poly-articulée pour la microscopie corrélative AFM-in-SEM. Un système robotique AFM à 6 ddls (3 translations et 3 rotations) est développé et intégré à l'intérieur d'un MEB. La capacité de contrôler 3 positions et 3 rotations d'une sonde AFM de taille micrométrique tout en maintenant le centre de rotation à proximité d'une micro-structure est un véritable défi. Cela est principalement dû aux incertitudes inhérentes à l'assemblage des systèmes micro-robotiques et aux jeux mécaniques dans les articulations du robot qui sont du même ordre de grandeur que la précision requise pour le positionnement de la sonde AFM. Les méthodes d'étalonnage des robots et la théorie du contrôle peuvent cependant surmonter ces limitations, comme le démontre cette thèse. Des stratégies de contrôle et une interface utilisateur sont étudiées pour faire fonctionner le système d'imagerie corrélative multi-ddl de manière polyvalente et intuitive. Plusieurs caractéristiques clés qui vont au-delà de l'état de l'art sont mises en œuvre, notamment - Le contrôle par vision électronique (MEB) permet l'atterrissage rapide et automatisé d'une sonde AFM sur un échantillon de taille micrométrique, avec une robustesse par rapport au grossissement du MEB. L'utilisateur peut sélectionner n'importe quelle région d'intérêt (ROI) sur un échantillon en cliquant simplement sur l'écran du MEB. Quel que soit le grossissement du MEB, l'algorithme de contrôle assure un atterrissage sûr de la sonde AFM sur la région d'intérêt. La surface de l'échantillon peut atteindre plusieurs centimètres carrés et le positionnement peut être réalisé avec une précision micrométrique. - Rotation dans le plan et hors du plan d'un échantillon par rapport à la sonde AFM tout en maintenant le centre de rotation autour de la pointe de l'AFM. Le centre de rotation est défini par l'utilisateur par un clic de souris sur l'écran du MEB. Cette fonction est utile pour les tâches de manipulation et de topographie, ainsi que pour les observations multi-angles d'un échantillon à l'intérieur d'un MEB. - Modes de sélection de la trajectoire et de la vitesse de la sonde AFM. Mode AFM à faible vitesse pour une imagerie topographique détaillée. Mode AFM rapide (4fps) pour des observations dynamiques à l'échelle nanométrique. Les utilisateurs ont également accès aux paramètres de contrôle. Ils peuvent être modifiés en fonction de leurs besoins. - Mode AFM mosaïque pour étendre la zone de balayage de la topographie à l'intérieur d'un MEB. Toutes ces caractéristiques s'appuient sur les travaux de recherche en robotique, mécatronique et contrôle réalisés au cours de la thèse. Ces derniers ont le potentiel d'ouvrir la porte à une nouvelle ère de microscopes à force atomique poly-articulés utilisés en microscopie corrélative<br>Correlative microscopy is the result of the combination of two or more microscopy techniques to provide complementary information on a sample. When using a scanning electron microscope (SEM) and an atomic force microscope (AFM), AFM-in-SEM correlative microscopy not only enables the 3D characterization of samples observed inside a SEM, but also the manipulation of micro- and nanostructures with an extremely high precision. This technique can be applied to various samples in biology, electronics and materials science. Although existing AFM-in-SEM solutions in the current state of the art are powerful, they require expert users; they are not versatile enough to be used for different types of tasks; and they use Cartesian AFM robots that severely limit the dexterity and performance of the imaging system. The aim of this thesis is to study and experiment an original concept of an AFM based on poly- articulated robotics for AFM-in-SEM correlative microscopy. A homemade 6 DoF (3 translations and 3 rotations) robotic AFM system is developed and integrated inside a SEM. The ability to control 3 positions and 3 rotations of a micrometer sized AFM probe while keeping the center of rotation at the close proximity of a micro-structure is very challenging. This is mainly due to the uncertainties inherent to the assembly of micro-robotic systems and clearances in the joints of the robot that are of the same order of magnitude as the required AFM probe positioning accuracy. Robot calibration methods and control theory can however overcome these limitations as demonstrated in the thesis. Control strategies and a user interface are studied to operate the multi DoF correlative imaging system in a versatile and intuitive way for low-level end users while keeping it enough powerful for high-level end users. Several key features that go beyond the state of the art are implemented, including - Vision based control for fast and automated landing of an AFM probe on a micrometer sized sample with robustness with respect to the SEM magnification. The user can select any region of interest (ROI) on a sample by simply performing a mouse click on the SEM screen. Whatever the SEM magnification, the control algorithm ensures a safe landing of the AFM probe on the ROI. The surface of the sample can be as high as several square centimeters and the positioning can be achieved with a micrometric precision. - In-plane and out-of-plane rotation of a sample relatively to the AFM probe while keeping the center of rotation around the tip of the AFM. The center of rotation is defined by the user with a mouse click on the SEM screen. This feature is useful for manipulation and topography tasks, as well as for multi-angle observations of a sample inside a SEM. - Trajectory/speed selection modes. Low speed AFM mode for a detailed topography imaging. Fast AFM mode (4fps) for dynamic observations at the nanoscale. The users also have access to the control parameters. They can be modified to suit their needs. - Mosaic AFM mode to extend the topography scanning area inside a SEM. All these features rely on research works in robotics, mechatronics and control made during the thesis. The latter has the potential to opens the door to a new era of poly-articulated atomic force microscopes used in correlative microscopy
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Hackley, Jason. "A Liquid-Helium-Free High-Stability Cryogenic Scanning Tunneling Microscope for Atomic-Scale Spectroscopy." Thesis, University of Oregon, 2015. http://hdl.handle.net/1794/19211.
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This dissertation provides a brief introduction into scanning tunneling microscopy, and then Chapter III reports on the design and operation of a cryogenic ultra-high vacuum scanning tunneling microscope (STM) coupled to a closed-cycle cryostat (CCC). The STM is thermally linked to the CCC through helium exchange gas confined inside a volume enclosed by highly flexible rubber bellows. The STM is thus mechanically decoupled from the CCC, which results in a significant reduction of the mechanical noise transferred from the CCC to the STM. Noise analysis of the tunneling current shows current fluctuations up to 4% of the total current, which translates into tip-sample distance variations of up to 1.5 picometers. This noise level is sufficiently low for atomic-resolution imaging of a wide variety of surfaces. To demonstrate this, atomic-resolution images of Au(111) and NaCl(100)/Au(111) surfaces, as well as of carbon nanotubes deposited on Au(111), were obtained. Other performance characteristics such as thermal drift analysis and a cool-down analysis are reported. Scanning tunneling spectroscopy (STS) measurements based on the lock-in technique were also carried out and showed no detectable presence of noise from the CCC. These results demonstrate that the constructed CCC-coupled STM is a highly stable instrument capable of highly detailed spectroscopic investigations of materials and surfaces at the atomic-scale.
A study of electron transport in single-walled carbon nanotubes (SWCNTs) was also conducted. In Chapter IV, STS is used to study the quantum-confined electronic states in SWCNTs deposited on the Au(111) surface. The STS spectra show the vibrational overtones which suggest rippling distortion and dimerization of carbon atoms on the SWCNT surface. This study experimentally connects the properties of well-defined localized electronic states to the properties of their associated vibronic states.
In Chapter V, a study of PbS nanocrystals was conducted to study the effect of localized sub-bandgap states associated with surface imperfections. A correlation between their properties and the atomic-scale structure of chemical imperfections responsible for their appearance was established to understand the nature of such surface states.
This dissertation includes both previously published/unpublished and co-authored material.
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Mayerich, David Matthew. "Acquisition and reconstruction of brain tissue using knife-edge scanning microscopy." Texas A&M University, 2003. http://hdl.handle.net/1969.1/563.
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A fast method for gathering large-scale data sets through the serial sectioning of
brain tissue is described. These data sets are retrieved using knife-edge scanning
microscopy, a new technique developed in the Brain Networks Laboratory at Texas
A&M University. This technique allows the imaging of tissue as it is cut by an
ultramicrotome.
In this thesis the development of a knife-edge scanner is discussed as well as the
scanning techniques used to retrieve high-resolution data sets. Problems in knife-edge
scanning microscopy, such as illumination, knife chatter, and focusing are discussed.
Techniques are also shown to reduce these problems so that serial sections of tissue can
be sampled at resolutions that are high enough to allow reconstruction of neurons at the
cellular level.
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Joseph, Serge. "Industrialisation d'un prototype de microscope acoustique." Valenciennes, 1989. https://ged.uphf.fr/nuxeo/site/esupversions/b8833446-48ed-4b35-8b84-a97acf200619.
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Eigenbrot, Ilya Vladimirovich. "A time-resolved confocal fluorescence microscope." Thesis, Imperial College London, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.342331.
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Lawrence, Andrew James. "Development of a Hybrid Atomic Force and Scanning Magneto-Optic Kerr Effect Microscope for Investigation of Magnetic Domains." PDXScholar, 2011. https://pdxscholar.library.pdx.edu/open_access_etds/147.
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We present the development of a far-field magneto-optical Kerr effect microscope. An inverted optical microscope was constructed to accommodate Kerr imaging and atomic force microscopy. In Kerr microscopy, magnetic structure is investigated by measuring the polarization rotation of light reflected from a sample in the presence of a magnetic field. Atomic force microscopy makes use of a probe which is scanned over a sample surface to map the topography. The design was created virtually in SolidWorks, a three-dimensional computer-aided drafting environment, to ensure compatibility and function of the various components, both commercial and custom-machined, required for the operation of this instrument. The various aspects of the microscope are controlled by custom circuitry and a field programmable gate array data acquisition card at the direction of the control code written in National Instrument LabVIEW. The microscope has proven effective for both Kerr and atomic force microscopy. Kerr images are presented which reveal the bit structure of magneto-optical disks, as are atomic force micrographs of an AFM calibration grid. Also discussed is the future direction of this project, which entails improving the resolution of the instrument beyond the diffraction limit through near-field optical techniques. Preliminary work on fiber probe designs is presented along with probe fabrication work and the system modifications necessary to utilize such probes.
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López, Ayón Gabriela. "Applying a commercial atomic force microscope for scanning near-field optical microscopy techniques and investigation of Cell-cell signalling." Thesis, McGill University, 2010. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=92400.
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The field of research of this thesis is Condensed Matter Physics applied to Biology. Specifically it describes the development of different Atomic Force Microscopy techniques and tools towards the study of living cells in physiological solution. Particular interest is put into the understanding of the influence of noise in the determination of ordered liquid layers above a mica surface - as work towards the study of the role of water and ions in biological processes - and the influence of "diving bell" to boost the Q factor and allow stable imaging and force spectroscopy with tips based on Scanning Near-field Optical Microscopy [LeDue, 2010 and LeDue, 2008]. By combining SNOM techniques as a local illumination method (and thus avoiding photo bleaching of individual molecules) and high resolution AFM techniques we will be able to investigate mechano-transduction and associated signaling in living cells and individual proteins.<br>Le domaine de recherche de cette thèse consiste en l'application de la physique de la matière condensée à la biologie. Plus précisément, ce travail décrit le développement de différentes techniques de Microscopie à Force Atomique (MFA) et d'outils permettant l'étude de cellules vivantes en solution physiologique. Un intérêt particulier est porté à la compréhension de l'influence du bruit dans la détermination de couches liquides ordonnées au-dessus d'une surface de mica - en tant que travail préalable à l'étude du rôle de l'eau et des ions dans les processus biologiques - et de l'influence d'une "cloche de plongée" pour renforcer le facteur Q ainsi que pour permettre l'imagerie stable et la spectrométrie de force avec des sondes basées sur la Microscopie Optique en Champ Proche (MOCP). En combinant des techniques MOCP, utilisées comme méthode d'éclairement local (évitant ainsi le photoblanchiment des molécules individuelles), et des techniques MFA haute résolution, nous serons capables d'investir la mécano-transduction et le signalement associé dans des cellules vivantes et dans des protéines individuelles.
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Clarke, Richard John. "Hydrodynamics of the atomic force microscope." Thesis, University of Nottingham, 2005. http://eprints.nottingham.ac.uk/10649/.
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With a proven ability to uncover fundamental biological processes, the atomic force microscope (AFM) represents one of the most valuable and versatile tools available to the biophysical sciences. We study the unsteady small-scale flows generated within the AFM by its sensing probe (a long thin cantilever), which have received relatively little attention to date, yet which are increasingly relevant in an age of microdevices. The early parts of this thesis investigate some canonical two-dimensional flows driven by oscillations of an infinite-length rigid cantilever. These prove amenable to analysis and enable us to investigate many of the important physical phenomena and compile a comprehensive collection of asymptotic expressions for the drag. The corresponding results lay out the influence of a nearby wall, geometry and oscillation frequency. The limitations of a two-dimensional approach are then explored through the development of a novel unsteady slender-body theory (USBT) for finite-length cylinders, an asymptotic treatment of which offers corrections to traditional resistive-force-theory (RFT) methods by accounting for geometric factors and flow inertia. These ideas are then extended to the study of thin rectangular plates. Two key parameters are identified which promote two-dimensionality in the flow, namely the frequency of oscillation and the proximity of a nearby boundary. We then examine flexible cylinders and plates by coupling the hydrodynamics to linearized elastic beam and plate equations, which simulate the hydrodynamically-damped high-speed deformable motion of the AFM's cantilever, when driven either externally or by Brownian motion. In the latter case, we adopt an approach which offers notable improvements over the most advanced method currently available to the AFM community.
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Nowak, Derek Brant. "The Design of a Novel Tip Enhanced Near-field Scanning Probe Microscope for Ultra-High Resolution Optical Imaging." PDXScholar, 2010. https://pdxscholar.library.pdx.edu/open_access_etds/361.
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Traditional light microscopy suffers from the diffraction limit, which limits the spatial resolution to λ/2. The current trend in optical microscopy is the development of techniques to bypass the diffraction limit. Resolutions below 40 nm will make it possible to probe biological systems by imaging the interactions between single molecules and cell membranes. These resolutions will allow for the development of improved drug delivery mechanisms by increasing our understanding of how chemical communication within a cell occurs. The materials sciences would also benefit from these high resolutions. Nanomaterials can be analyzed with Raman spectroscopy for molecular and atomic bond information, or with fluorescence response to determine bulk optical properties with tens of nanometer resolution. Near-field optical microscopy is one of the current techniques, which allows for imaging at resolutions beyond the diffraction limit. Using a combination of a shear force microscope (SFM) and an inverted optical microscope, spectroscopic resolutions below 20 nm have been demonstrated. One technique, in particular, has been named tip enhanced near-field optical microscopy (TENOM). The key to this technique is the use of solid metal probes, which are illuminated in the far field by the excitation wavelength of interest. These probes are custom-designed using finite difference time domain (FDTD) modeling techniques, then fabricated with the use of a focused ion beam (FIB) microscope. The measure of the quality of probe design is based directly on the field enhancement obtainable. The greater the field enhancement of the probe, the more the ratio of near-field to far-field background contribution will increase. The elimination of the far-field signal by a decrease of illumination power will provide the best signal-to-noise ratio in the near-field images. Furthermore, a design that facilitates the delocalization of the near-field imaging from the far-field will be beneficial. Developed is a novel microscope design that employs two-photon non-linear excitation to allow the imaging of the fluorescence from almost any visible fluorophore at resolutions below 30 nm without changing filters or excitation wavelength. The ability of the microscope to image samples at atmospheric pressure, room temperature, and in solution makes it a very promising tool for the biological and materials science communities. The microscope demonstrates the ability to image topographical, optical, and electronic state information for single-molecule identification. A single computer, simple custom control circuits, field programmable gate array (FPGA) data acquisition, and a simplified custom optical system controls the microscope are thoroughly outlined and documented. This versatility enables the end user to custom-design experiments from confocal far-field single molecule imaging to high resolution scanning probe microscopy imaging. Presented are the current capabilities of the microscope, most importantly, high-resolution near-field images of J-aggregates with PIC dye. Single molecules of Rhodamine 6G dye and quantum dots imaged in the far-field are presented to demonstrate the sensitivity of the microscope. A comparison is made with the use of a mode-locked 50 fs pulsed laser source verses a continuous wave laser source on single molecules and J-aggregates in the near-field and far-field. Integration of an intensified CCD camera with a high-resolution monochromator allows for spectral information about the sample. The system will be disseminated as an open system design.
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Ratcliff, Marc. "Europe and the microscope in the Enlightenment." Thesis, University College London (University of London), 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.271134.
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Guénon, Stefan [Verfasser], and Reinhold [Akademischer Betreuer] Kleiner. "Cryogenic Scanning Laser Microscopy : Investigation of large BSCCO mesas and development of a polarizing microscope / Stefan Guénon ; Betreuer: Reinhold Kleiner." Tübingen : Universitätsbibliothek Tübingen, 2011. http://d-nb.info/1162627085/34.
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Marturi, Naresh. "Vison and visual servoing for nanomanipulation and nanocharacterization using scanning electron microscope." Thesis, Besançon, 2013. http://www.theses.fr/2013BESA2014/document.
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Avec les dernières avancées en matière de nanotechnologies, il est devenu possible de concevoir, avec une grande efficacité, de nouveaux dispositifs et systèmes nanométriques. Il en résulte la nécessité de développer des méthodes de pointe fiables pour la nano manipulation et la nano caractérisation. La d´étection directe par l’homme n’ étant pas une option envisageable à cette échelle, les tâches sont habituellement effectuées par un opérateur humain expert `a l’aide de microscope électronique à balayage équipé de dispositifs micro nano robotiques. Toutefois, en raison de l’absence de méthodes efficaces, ces tâches sont toujours difficiles et souvent fastidieuses à réaliser. Grâce à ce travail, nous montrons que ce problème peut être résolu efficacement jusqu’ à une certaine mesure en utilisant les informations extraites des images. Le travail porte sur l’utilisation des images électroniques pour développer des méthodes automatiques fiables permettant d’effectuer des tâches de nano manipulation et nano caractérisation précises et efficaces. En premier lieu, puisque l’imagerie électronique à balayage est affectée par les instabilités de la colonne électronique, des méthodes fonctionnant en temps réel pour surveiller la qualité des images et compenser leur distorsion dynamique ont été développées. Ensuite des lois d’asservissement visuel ont été développées pour résoudre deux problèmes. La mise au point automatique utilisant l’asservissement visuel, développée, assure une netteté constante tout au long des processus. Elle a permis d’estimer la profondeur inter-objet, habituellement très difficile à calculer dans un microscope électronique à balayage. Deux schémas d’asservissement visuel ont été développés pour le problème du nano positionnement dans un microscope électronique. Ils sont fondés sur l’utilisation directe des intensités des pixels et l’information spectrale, respectivement. Les précisions obtenues par les deux méthodes dans diff érentes conditions expérimentales ont été satisfaisantes. Le travail réalisé ouvre la voie à la réalisation d’applications précises et fiables telles que l’analyse topographique,le sondage de nanostructures ou l’extraction d’ échantillons pour microscope électronique en transmission<br>With the latest advances in nanotechnology, it became possible to design novel nanoscale devicesand systems with increasing efficiency. The consequence of this fact is an increase in the need for developing reliable and cutting edge processes for nanomanipulation and nanocharacterization. Since the human direct sensing is not a feasible option at this particular scale, the tasks are usually performedby an expert human operator using a scanning electron microscope (SEM) equipped withmicro-nanorobotic devices. However, due to the lack of effective processes, these tasks are always challenging and often tiresome to perform. Through this work we show that, this problem can be tackle deffectively up to an extent using the microscopic vision information. It is concerned about using the SEM vision to develop reliable automated methods in order to perform accurate and efficient nanomanipulation and nano characterization. Since, SEM imaging is affected by the non-linearities and instabilities present in the electron column, real time methods to monitor the imaging quality and to compensate the time varying distortion were developed. Later, these images were used in the development of visual servoing control laws. The developed visual servoing-based autofocusing method ensures a constant focus throughout the process and was used for estimating the inter-object depth that is highly challenging to compute using a SEM. Two visual servoing schemes were developed toperform accurate nanopositioning using a nanorobotic station positioned inside SEM. They are basedon the direct use of global pixel intensities and Fourier spectral information respectively. The positioning accuracies achieved by both the methods at different experimental conditions were satisfactory.The achieved results facilitate in developing accurate and reliable applications such as topographic analysis, nanoprobing and sample lift-out using SEM
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El, Khoury Diana. "Towards the use of electrostatic force microscopy to study interphases in nanodielectric materials." Thesis, Montpellier, 2017. http://www.theses.fr/2017MONTS078/document.
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Les interphases sont souvent considérées comme responsables des propriétés physiques des nanodiélectriques inexplicables par les lois de mélange. La prédiction de la permittivité diélectrique des nanodiélectriques nécessite de reconsidérer la permittivité intrinsèque et le volume de l'interphase. Malgré le besoin d'une caractérisation locale de ces régions interfaciales nanométriques, aucune méthode expérimentale fiable n'a encore été développée. La Microscopie à Force Electrostatique (EFM) constitue une technique prometteuse pour atteindre ce but. L'objectif de cette thèse est de développer des protocoles expérimentaux et des méthodes d’interprétations du signal appropriés pour évaluer l’aptitude de l’EFM à l'étude des interphases dans les nanodiélectriques. Nous avons eu recours d’abord à des simulations numériques par éléments-finis pour approfondir notre compréhension de l'interaction entre une sonde EFM et plusieurs types d'échantillons nanostructurés, permettant par la suite de simuler la réponse spécifique face à un nanocomposite possédant une interphase. Nous avons proposé un modèle électrostatique de nanodiélectrique possédant trois phases, selon lequel, nous avons conçu et synthétisé des échantillons modèles aux propriétés connues afin de jouer le rôle de matériaux nanodiélectriques de référence pour les mesures EFM. Par conséquent, nous avons pu développer plusieurs protocoles expérimentaux et d’analyses du signal utilisant des modes DC et AC de détection du gradient de force pour caractériser les interphases dans des nanocomposites. Ces techniques constituent un ensemble polyvalent de méthodes d’étude des interphases avec un impact réduit des effets parasites communément convolués dans les signaux EFM. Enfin, une quantification de la permittivité de l'interphase de nos échantillons modèles a été possible par corrélation avec nos simulations numériques<br>Interphases are usually considered to be responsible for the physical properties of nanodielectrics unexplainable by general mixture laws. The prediction of the effective dielectric permittivity of these materials needs to reconsider the intrinsic permittivity and the volume of the interphase. Despite the urge for a local characterization of these nanometric interfacial regions, no reliable experimental method has been developed yet. The Electrostatic Force Microscope (EFM) constitutes a promising technique to fulfill this objective. The aim of this thesis is to develop appropriate experimental protocols and signal analysis to explore the ability of EFM to the study of interphases in nanodielectrics. We first resorted to finite-element numerical simulations in order to deeper our understanding of the interaction between an EFM probe and several types of nanostructured samples, allowing to simulate afterwards the specific response to a nanocomposite possessing an interphase. We proposed a three-phase electrostatic model of a nanodielectric, upon which, we designed and synthesized model samples of known properties to play the role of a reference nanodielectric material for EFM measurements. Consequently, we were able to develop several experimental protocols and signal analysis with both DC and AC force gradient EFM modes. These techniques offer versatile methods to characterize interphases with reduced impact of the parasitic effects commonly convoluted within EFM signals. Finally, a quantification of the interphase in our nanodielectric model samples was possible thanks to correlation with our numerical simulations
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Lhenry, Sébastien. "Nouveaux développements en microscopie électrochimique (SECM) pour l'analyse de surfaces fonctionnalisées." Thesis, Rennes 1, 2014. http://www.theses.fr/2014REN1S150/document.
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Ce manuscrit décrit les travaux menés pour le développement et l'utilisation du microscope électrochimique (SECM) pour l'étude des surfaces fonctionnelles. Cette microscopie à sonde locale, utilisant les propriétés des ultramicroélectrodes (UMEs), permet l'analyse de la réactivité chimique à la surface d'un échantillon. Le but de ces travaux est alors l'analyse de surfaces fonctionnalisées par une couche organique. Dans un premier temps, nous avons développé la méthodologie du SECM par l'utilisation de sonde redox spécifique. D'abord, cela nous a permis de déterminer la distance entre l'électrode-sonde et la surface grâce à un médiateur redox irréversible comme les polyaromatiques. Ensuite, nous avons pu observer des propriétés physico-chimiques d'une surface, notamment afin de différencier les différents modes de transport électronique au travers une couche organique, grâce à l'utilisation des catéchols ou des dendrimères redox. Nous avons également commencé le développement d'un nouveau mode d'utilisation du SECM : le SECM transitoire. Dans un deuxième temps, nous avons utilisé le SECM afin d'analyser trois surfaces fonctionnalisées. La première surface est un substrat d'or fonctionnalisé par des calixarènes. La deuxième est une surface de verre sur laquelle il a été déposée des nanoparticules photosensibles, appelées « quantum rods ». Et la dernière est un substrat de silicium modifié par un récepteur anthracène. Finalement, la dernière étude qui a été menée concerne l'écriture et la lecture d'une monocouche organique grâce à la sonde du SECM. Ces modifications localisées de la couche organique sont alors de la taille de l'UME, soit de l'ordre du micromètre. Dans notre cas nous avons choisi la modification de monocouche organique auto-assemblée (SAM) sur un substrat d'or grâce à la réaction de couplage de Huisgen. En plus, de l'amélioration de nos conditions de fonctionnalisation, nous avons pu mettre en évidence un phénomène de propagation catalytique en surface grâce à un complexe spécial de cuivre (II) contenant une fonction éthynyle<br>This manuscript describes the work done for the development and use of electrochemical microscope (SECM) for the investigations of functional surfaces. This local probe microscopy, using the properties of ultramicroelectrodes (UMEs), permits the analysis of chemical reactivity on the sample’s surface. The aim of this work is then analyzing surfaces functionalized with an organic layer. Initially, we have developed the SECM methodology by using specific redox probes. First, it allowed us to determine the absolute distance between the electrode and the surface, thanks to an irreversible redox mediator such as polyaromatic. Then, we could observe the physicochemical surface properties, in particular to differentiate the electronic transfer modes through an organic layer, by using catechols or redox dendrimers. We have also started the development of a new mode of SECM: transient SECM. Secondly, we used the SECM to analyze three functionalized surfaces. The first surface is a gold substrate functionalized by calixarenes. The second is a glass side on which it was deposed photosensitive nanoparticles, called "quantum rods" and the last sample is a substrate of silicon modified by anthracene receptors. Finally, the last study concerns the writing and reading on an organic monolayer with the SECM probe. These localized changes of the organic layer are about the size of the UME, of the order of a micrometer. In our case, we chose the modification of self-assembled organic monolayer (SAM) on a gold substrate by the Huisgen coupling reaction. In addition to the improvement of our conditions of functionalization, we were able to demonstrate a phenomenon of surface catalytic propagation due to a special complex of copper (II) containing ethynyl functions
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Zdankowski, Piotr. "Adaptive optics stimulated emission depletion microscope for thick sample imaging." Thesis, University of Dundee, 2018. https://discovery.dundee.ac.uk/en/studentTheses/90e27151-f51c-4c12-b9dd-2bc78beb2321.
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Over the past few decades, fluorescence microscopy has proven to become the most widely used imaging technique in the field of life sciences. Unfortunately, all classical optical microscopy techniques have one thing in common: their resolution is limited by the diffraction. Thankfully, due to the very strong interest, development of fluorescent microscopy techniques is very intense, with novel solutions surfacing repeatedly. The major breakthrough came with the appearance of super-resolution microscopy techniques, enabling imaging well below the diffraction barrier and opening the new era of nanoscopy. Among the fluorescent super-resolution techniques, Stimulated Emission Depletion (STED) microscopy has been particularly interesting, as it is a purely optical technique which does not require post image processing. STED microscopy has proven to resolve structures down to the molecular resolution. However, super-resolution microscopy is not a cure to all the problems and it also has its limits. What has shown to be particularly challenging, was the super-resolution imaging of thick samples. With increased thickness of biological structures, the aberrations increase and signal-to-noise (SNR) decreases. This becomes even more evident in the super-resolution imaging, as the nanoscopic techniques are especially sensitive to aberrations and low SNR. The aim of this work is to propose and develop a 3D STED microscope that can successfully image thick biological samples with nanoscopic resolution. In order to achieve that, adaptive optics (AO) has been employed for correcting the aberrations, using the indirect wavefront sensing approach. This thesis presents a custom built 3D STED microscope with the AO correction and the resulting images of thick samples with resolution beyond diffraction barrier. The developed STED microscope achieved the resolution of 60nm in lateral and 160nm in axial direction. What is more, it enabled super-resolution imaging of thick, aberrating samples. HeLa, RPE-1 cells and dopaminergic neuron differentiated from human IPS cells were imaged using the microscope. The results shown in this thesis present 3D STED imaging of thick biological samples and, what is particularly worth to highlight, 3D STED imaging at the 80μm depth, where the excitation and depletion beams have to propagate through the thick layer of tissue. 3D STED images at such depth has not been reported up to date.
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Haenssler, Olaf Christian. "Multimodal sensing and imaging technology by integrated scanning electron, force, and near-field microwave microscopy and its application to submicrometer studies." Thesis, Lille, 2018. http://www.theses.fr/2018LIL1I006.
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La combinaison de plusieurs procédés d’imagerie et de mesure permet d’obtenir des ensembles de données complémentaires et parfois uniques. A l’aide d’une technique hybride de microscopie présentant des modalités de mesure différentes et des enregistrements synchrones, on peut recueillir des informations complémentaires sur des échantillons à l’échelle nanométrique. De plus, l’intégration de procédés nanorobotiques et de logiciels open-source permet une approche technologique pour la recherche sur les semi-conducteurs et les sciences des matériaux. Ce travail démontre le potentiel d’une telle technologie. Ce démonstrateur fonctionne dans la chambre d‘un MEB et sert de plateforme technologique dans laquelle sont intégrés différentes modalités, technologies et procédés. Un AFM basé sur un interféromètre optique compact permet l’imagerie de la topographie de surface tandis qu’un microscope à micro-ondes à balayage enregistre les caractéristiques électromagnétiques dans la gamme de fréquence des micro-ondes, le tout opérant dans le même MEB. L’engin est contrôlé par un ensemble de logiciels qui est optimisé pour la nanorobotique basée sur l‘imagerie. Ce démonstrateur technologique permet d’observer en direct la région d’intérêt à l’aide du microscope électronique tandis qu’est effectuée en champ proche la caractérisation de la surface de l’échantillon par intermédiaire des micro-ondes évanescentes et des forces intermoléculaires. Ensuite, est présenté un standard multimodal de test et qui valide la fonctionnalité de l’instrument démonstrateur. Le présent travail est complété par une analyse électrique de capacités MOS ainsi que leur approximation destinée au calibrage<br>Various disciplines of micro- and nanotechnology requires combinatorial tools for the investigation, manipulation and transport of materials in the submicrometer range. The coupling of multiple sensing and imaging techniques allows for obtaining complementary and often unique datasets of samples under test. By means of an integrated microscopy technique with different modalities, it is possible to gain multiple information about nanoscale samples by recording at the same time. The expansion with nanorobotics and an open-source software framework, leads to a technology approach for semiconductor research and material science. This work shows the potential of such a multimodal technology approach by focusing on a demonstrator setup. It operates under high-vacuum conditions inside the chamber of a Scanning Electron Microscope and serves as a technology platform by fusing various microscopy modalities, techniques and processes. An Atomic Force Microscope based on a compact, optical interferometer performs imaging of surface topography, and a Scanning Microwave Microscope records electromagnetic properties in the microwave frequency domain, both operating inside an SEM. A software framework controls the instrument. The setup allows for observing with SEM, while imaging and characterizing with interacting evanescent microwaves and intermolecular forces simultaneously. In addition, a multimodal test standard is introduced and subsequently confirms the functionality of the demonstrator. Within this context, the work also includes an electrical analysis of micro-scale MOS capacitors, including an approximation for use in the calibration
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Girirajan, Thanu Prabha Kalambur. "Development of Spectral Imaging Microscope for Single Molecule Studies in Complex Biological Systems." Fogler Library, University of Maine, 2007. http://www.library.umaine.edu/theses/pdf/GirirajanTPK2007.pdf.
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Chang, Michael Ming Yuen. "A computer-controlled system in transmission electron microscopy." Thesis, University of Cambridge, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.292941.
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Fontes, Adriana. "Sistema de micromanipulação e microanálise com pinças óticas." [s.n.], 2004. http://repositorio.unicamp.br/jspui/handle/REPOSIP/277500.
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Orientador: Carlos Lenz Cesar<br>Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Fisica "Gleb Wataghin"<br>Made available in DSpace on 2018-08-04T14:59:07Z (GMT). No. of bitstreams: 1
Fontes_Adriana_D.pdf: 3876787 bytes, checksum: 3edd1575e993d4685d7e82c8a3f77cfb (MD5)
Previous issue date: 2004<br>Resumo: Não informado<br>Abstract: Not informed.<br>Doutorado<br>Física<br>Doutor em Ciências
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Bredekamp, Adriaan Hendrik. "New feedback control for a scanning tunneling microscope." Thesis, Cape Technikon, 1999. http://hdl.handle.net/20.500.11838/1129.
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Thesis (MTech(Electrical Engineering))--Cape Technikon, Cape Town,1999<br>This thesis describes the design and implementation of a new feedback controller for a
scanning tunneling microscope or STM. The previous controller had several shortcomings
when it came to the data throughput rate of the data acquisition system, the scan rate, and the
way the data was stored and displayed.
The initial investigation was done to determine the most cost effective way to implement the
data acquisition system. Various approaches such as DSP systems, analogue systems and
microcontroller systems were looked at. The investigation also looked at the best way to get
the data from the Z directional control loop to the PC for displaying the data. The final choice
was to use an ultra fast microcontroller for the control loop implementation and to change the
DOS based software for Windows based software.
The embedded system was divided into two parts. The first was the controller for the X and
Y scan directions, and the second was for the Z scan direction. A digital PI control loop was
implemented on the Z controller to control the height of the scan tip above the specimen
surface. The microcontroller that was chosen for this was the Microchip PIC17c43. The data
transfer to the PC was done with a PC-14 programmable digital input/output card. Two
options for the implementation of the PC-14 software were considered. The first option was
the software that was bundled with the card. This software proved to be very slow, so special
device-driver-based software was developed to control the PC-14 card and the data transfer to
and from the Pc. The PC software was implemented using Visual C++.
Both the XY and the Z controllers proved to be working satisfactorily in the existing STM
arrangement. It was discovered that the XY controller was overloaded with the many tasks
that it has to perform, and a suitable alternative system to replace the XY controller is
proposed. The selection of the PC that will be used for the data acquisition system is also
discussed. It was found that this choice had a very big influence on the design of the final
system because of the difference in PC bus design. Several proposals to increase the
functionality of the PC software are also made.
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Clark, Spencer C. "Development of the Evanescent Wave Atomic Force Microscope." Diss., Virginia Tech, 2005. http://hdl.handle.net/10919/29704.
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The conventional atomic force microscope (AFM) is equipped with a single optical detection system. Probe-sample separation is determined in an independent deflection with respect to AFM z-translation experiment. This method of determining probe-surface separation is relative, susceptible to drift and does not provide real time separation information. The evanescent wave atomic force microscope (EW-AFM) utilizes a second, independent detection system to determine absolute probe-surface separation in real time. The EW-AFM can simultaneously acquire real-time force and probe-sample separation information using the optical lever and evanescent scattering detection systems, respectively. The EW-AFM may be configured with feedback on the optical-lever system for constant force applications or with feedback on evanescent wave scattering intensity for constant height applications.
Scattering of the evanescent wave exponential decay profile is used to determine probe-surface separation. Sub-micron sized dielectric and metallic probes show exponential scattering profiles, micron sized polystyrene and borosilicate microspheres show non-exponential profiles when they are affixed beneath the cantilever tip. By affixing the microspheres to the end of the AFM cantilever exponential and non-exponential profiles were observed.
The EW-AFM can be used to conduct force-distance and imaging experiments. The EW-AFM was used to measure the thickness of surfactant bilayers formed at the silica-solution interface using silicon nitride AFM tips. The presence of a refractive index difference between the surfactant bilayer and the solution does not influence the accuracy of the surfactant bilayer thickness measurement. The EW-AFM was used to scan a 2 x 2 micron area in constant height mode. The probe was brought to within 6 nanometers of a planar dielectric surface using the evanescent wave intensity as a height reference with accuracy of ± 1 nm. This capability may be utilized to observe charge heterogeneity at the solid-liquid interface with nanometer lateral resolution or to map chemical functional group heterogeneity based on perturbations to the electrical double layer.
The EW-AFM evanescent scattering system has an absolute separation resolution of 0.3 nm compared to 1.0 nm relative separation resolution for the optical lever system. In constant scattering (constant height) mode the real time separation precision is about 2 nm.<br>Ph. D.
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Humphry, Martin James. "Novel scanning probe microscope instrumentation with applications in nanotechnology." Thesis, University of Nottingham, 2000. http://eprints.nottingham.ac.uk/13743/.
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A versatile scanning probe microscope controller has been constructed. Its suitability for the control of a range of different scanning probe microscope heads has been demonstrated. These include an ultra high vacuum scanning tunnelling microscope, with which atomic resolution images of Si surfaces was obtained, a custom-built atomic force microscope, and a custom-built photon emission scanning tunnelling microscope. The controller has been designed specifically to facilitate data acquisition during molecular manipulation experiments. Using the controller, the fullerene molecule C60 has been successfully manipulated on Si(100)-2x1 surfaces and detailed data has been acquired during the manipulation process. Evidence for two distinct modes of manipulation have been observed. A repulsive mode with success rates up to 90% was found to occur with tunnel gap impedances below 2GΩ, while between 2GΩ and 8GΩ attractive manipulation events were observed, with a maximum success rate of ~8%. It was also found that the step size between feedback updates had a significant effect on tip stability, and that dwell time of the STM tip at each data point had a critical effect on manipulation probability. A multi-function scanning probe microscope head has been developed capable of operation as a scanning tunnelling microscope and an atomic force microscope in vacuum and a magnetic field of 7T. The custom-built controller also presented here was used to control the head. A three-axis inertial sliding motor was developed for the head, capable of reproducible step sizes of <1000Å. In addition, an optical fibre interferometer was constructed with a sensitivity of 0.2Å/ √Hz. Preliminary development of a magnetic resonance force microscope mode has also been performed, with initial results showing such a system to be feasible.
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Reagan, Michael A. "Electric Charging and Nanostructure Formation in Polymeric Films using Combined Amplitude-Modulated Atomic Force Microscopy Assisted Electrostaitc Nanolithography and Electric Force Microscopy." University of Akron / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=akron1239637956.
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Jarasch, Markus. "Interfacing a Computer to a Scanning Tunneling Microscope." PDXScholar, 1994. https://pdxscholar.library.pdx.edu/open_access_etds/5047.
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A program was written in 'C' to control the functions of an already existing Scanning Tunneling Microscope (STM). A DAS-1601 data acquisition card (from Keithley Data Acquisition) was installed together with its drivers for 'C' on a computer with a 486-DX motherboard. The computer was interfaced to the electronics of the STM. Images taken of HOPG (highly oriented pyrolitic graphite) were of a reasonable quality and showed atomic resolution.
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Le, Cunuder Anne. "Étude expérimentale des forces de Casimir." Thesis, Lyon, 2017. http://www.theses.fr/2017LYSEN007/document.
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L'étude des fluctuations dans les milieux confinés constitue un domaine de recherche très récent, que ce soit du point de vue théorique ou expérimental. Afin d'analyser le rôle du confinement sur les propriétés des fluctuations de densité dans un mélange binaire, nous avons développé un système de mesure d'une grande sensibilité, où l'intensité des fluctuations et leur longueur de corrélation peuvent être amplifiées. L'idée consiste à travailler proche du point critique d'une transition de phase de démixion d'un mélange binaire. En effet, la longueur de corrélation augmente exponentiellement lorsqu'on s'approche de la température Tc du point critique de démixion.Nous avons développé un montage permettant de confiner le mélange entre un échantillon plan et une sphère colloïdale attachée à l'extrémité d'un levier de Microscope à Force Atomique (AFM). D'après les prédictions de Fisher et De Gennes, un effet intéressant émerge lorsque la longueur de corrélation est comparable avec la taille du confinement: les deux surfaces vont soit s'attirer, soit se repousser suivant les préférences d'adsorption des composants du mélange pour chacune des surfaces. On nomme cet effet l'effet Casimir critique, en référence à la force de Casimir électrodynamique qui résulte du confinement des fluctuations quantiques du champ électromagnétique.Durant cette thèse, nous avons mesuré la force de Casimir électrodynamique avec le système de mesure que nous avons développé, d'abord dans une atmosphère d'azote puis dans l'éthanol. Ces mesures prouvent que notre appareil de mesure est assez sensible pour mesurer des forces très faibles de l'ordre de la dizaine de pN. Les forces mesurées sont comparées à la théorie de Lifshitz, où les effets de conductivité finie des surfaces sont considérées<br>The study of density fluctuations inside confined liquid systems has received the attention of recent theoretical and experimental papers. In order to analyze the role of confinement on the statistical properties of fluctuations, we developed a highly sensitive system where the intensity of fluctuations, as well as their spatial correlation length can be simply tuned. The idea will be to enhance the role of fluctuations working close to the critical temperature Tc of a second order phase transition in a binary mixture. Indeed, the correlation length dramatically increases when one approaches the critical demixion point.The confinement is obtained by using a sphere-plane geometry with a colloidal particle attached to the cantilever of an Atomic Force Microscope (AFM). When the correlation length is comparable with the distance of confinement, Fisher and De Gennes predicted the existence of an interesting effect: the two surfaces will be submitted to either an attracting or a repelling force, depending on boundary conditions. This effect is called the critical Casimir force in reference to the quantum Casimir force resulting from the confinement of quantum fluctuations of the electromagnetic field.During this thesis, we measured the quantum Casimir force between the sphere and the plate, first in a nitrogen atmosphere and then in ethanol, showing that the developed instrument is sufficiently sensible to measure very weak force, of the same order of magnitude or even weaker than the critical Casimir force. Measurements are compared to Lifshitz theory, taking into account the finite conductivity of surfaces
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Löfgren, André. "Detection of electron vortex beams : Using a scanning transmission electron microscope." Thesis, Uppsala universitet, Materialteori, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-255330.
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Electron vortex beams (EVBs) are electron beams with a doughnut-like intensity profile, carrying orbital angular momentum due to their helical phase shift distribution. When employed in an electron microscope, they are expected to be efficient for the detection of magnetic signals. In this report I have investigated high angle annular dark field (HAADF) images obtained using EVBs. This was done for 300 K and 5K. For 5 K, I also compared HAADF images from an ordinary electron beam with HAADF images from an electron vortex beam. What was found was that EVBs produced doughnuts around the atomic columns. However, when taking the size of the electron source into account, this phenomena could no longer be observed. When comparing images from EVBs with images from ordinary electron beams, I found that the intensity of scattered electrons around atomic columns was broader for EVBs. This was persistent even after taking the source size into account.<br>Elektronvirvelstrålar (EVS) är elektronstrålar med en munk-liknande intensitetsprofil. Dessa bär på rörelsemängdsmoment på grund av sin fasdistribution. När de används i ett elektronmikroskop förväntas de vara effektiva för detektering av magnetiska signaler. I denna uppsats har jag undersökt high angle annular dark field (HAADF) bilder som erhållits med hjälp av EVS. Detta gjordes för 300 K och 5K. För 5 K, jämförde jag även HAADF bilder från en vanlig elektronstråle med HAADF bilder från en elektronvirvelstråle. Vad jag fann var att EVS producerade en munkformad intensitetsfördelning runt atomerna. Men när hänsyn till storleken på elektronkällan togs i beaktande kunde inte detta fenomen observeras längre. När bilder från EVS jämfördes med bilder från vanliga elektronstrålar, fann jag att intensiteten av spridda elektroner runt atomkolumnerna var bredare för EVS. Detta kunde observeras även efter att jag tagit hänsyn till elektronkällans storlek.
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Foland, Jed Rivera. "The body through the lens : anatomy and medical microscopy during the enlightenment." Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:3a82a1a2-15fd-458e-a566-6d52ed59d8b7.
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This thesis examines the role of microscope technology in informing medical and anatomical knowledge during the Enlightenment. Past historians have claimed that microscopy generally stagnated until the popularisation of achromatic microscopes and cell theory in the middle of the nineteenth century. As evidence for this decline, historians have pointed to the poor quality and slow development of microscope designs until the popularisation of achromatic microscopes in the 1820s. In contrast, this thesis highlights the role of specific Enlightenment-era microscopes in answering medical and anatomical questions. It suggests that medical microscopy was far more advanced than previous scholarship has ascertained. Thus far, instrument historians have focused more attention on competing instrument makers as opposed to rival instrument users. This thesis presents several case studies which explore both makers and users. These concern the histories of Enlightenment-era epidemiology, reproduction theory, anatomy, and physiology as well as the different types of microscopes which influenced these fields. In terms of methodology, this thesis neither follows nor casts doubt on any particular theory of historical development; rather, it attempts to shed further light on available primary sources and their contexts. Presenting key case studies illustrates the difficulties that early microscope users faced in acquiring and publishing new observations. To explore the practice of early microscopy further, this thesis presents re-enactments of these case studies using Enlightenment-era microscopes and modern tissue samples. Thus, this thesis is a call to broaden the scope of primary sources available to historians of science and medicine to include instruments and re-enactments. This thesis finds that technological advances did not correlate to microscopical discovery in medicine or anatomy. Both simple and complex microscope designs aided anatomical and medical research. Broader advances in anatomy, physiology, and medical etiology dictated the utility of medical microscopy. Although various groups, such as the French clinicians, saw little need for microscopy towards the end of the eighteenth century, microscope-based evidence continued to play a diagnostic role among lesser-known practitioners despite its lack of visibility in medical literature.
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Shen, Jie. "Construction, evaluation and application of a scanning tunnelling microscope." Thesis, De Montfort University, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.257605.
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Kozell, Monte Allen. "Investigation of the Acoustic Response of a Confined Mesoscopic Water Film Utilizing a Combined Atomic Force Microscope and Shear Force Microscope Technique." Thesis, Portland State University, 2018. http://pqdtopen.proquest.com/#viewpdf?dispub=10824892.
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<p> An atomic force microscopy beam-like cantilever is combined with an electrical tuning fork to form a shear force probe that is capable of generating an acoustic response from the mesoscopic water layer under ambient conditions while simultaneously monitoring force applied in the normal direction and the electrical response of the tuning fork shear force probe. Two shear force probes were designed and fabricated. A gallium ion beam was used to deposit carbon as a probe material. The carbon probe material was characterized using energy dispersive x-ray spectroscopy and scanning transmission electron microscopy. The probes were experimentally validated by demonstrating the ability to generate and observe acoustic response of the mesoscopic water layer.</p><p>
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Parshall, Daniel. "Phase imaging digital holography for biological microscopy." [Tampa, Fla.] : University of South Florida, 2004. http://purl.fcla.edu/fcla/etd/SFE0000285.
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Johnson, Lars. "Nanoindentation in situ a Transmission Electron Microscope." Thesis, Linköping University, Department of Physics, Chemistry and Biology, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-8333.
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<p>The technique of Nanoindentation <em>in situ</em> Transmission Electron Microscope has been implemented on a Philips CM20. Indentations have been performed on Si and Sapphire (<em>α-Al</em><em>2</em><em>O</em><em>3</em>) cut from wafers; Cr/Sc multilayers and <em>Ti</em><em>3</em><em>SiC</em><em>2</em> thin films. Different sample geometries and preparation methods have been evaluated. Both conventional ion and Focused Ion Beam milling were used, with different ways of protecting the sample during milling. Observations were made of bending and fracture of samples, dislocation nucleation and dislocation movement. Basal slip was observed upon unloading in Sapphire. Dislocation movement constricted along the basal planes were observed in <em>Ti</em><em>3</em><em>SiC</em><em>2</em>. Post indentation electron microscopy revealed kink formation in <em>Ti</em><em>3</em><em>SiC</em><em>2</em> and layer rotation and slip across layers in Cr/Sc multilayer stacks. Limitations of the technique are presented and discussed.</p>
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Shao, Yuhe. "Mems 3-D scan mirror for an endoscopic confocal microscope." Diss., Montana State University, 2005. http://etd.lib.montana.edu/etd/2005/shao/ShaoY1205.pdf.
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Şcheul, Ancuţa Teodora. "Two approaches for a simpler STED microscope using a dual-color laser or a single wavelength." Thesis, Grenoble, 2013. http://www.theses.fr/2013GRENY040/document.
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La microscopie STED (stimulated emission depletion ou déplétion par émission stimulée) est une des méthodes les plus répandues de microscopie de super-résolution. Dans un microscope STED, un faisceau en anneau se superpose avec le faisceau d'excitation et éteint les fluorophores en périphérie du faisceau d'excitation par émission stimulée. Au centre de l'anneau, où le faisceau STED a une intensité nulle, la fluorescence reste intacte. Cette technique nécessite un montage complexe dans lequel deux faisceaux laser, en général issus de deux sources différentes, doivent être parfaitement alignés et superposés. Dans ce travail de thèse, nous proposons deux configurations STED qui ont pour but de simplifier le montage et de réduire le coût total d'un tel système. L'idée de base dans les deux cas est d'utiliser la même source laser à la fois pour l'excitation et la déplétion par émission stimulée. Dans la première configuration, nous avons développé une source bicolore originale basée sur un laser Nd-YAG microchip. Ce laser microchip délivre simultanément des impulsions sub- ns à deux longueurs d'onde, 355 nm (excitation) et 532 nm (déplétion), qui sont générés par conversion harmonique à partir d'une émission laser Nd-YAG et offrent l'avantage d'être intrinsèquement alignées et synchronisées. Afin de trouver des colorants appropriés pour cette source particulière, nous avons développé une méthode de caractérisation et testé différents colorants Nous avons construit un microscope à partir de cette source laser et obtenu des images avec une résolution améliorée. La réduction du volume d'excitation a été confirmée par spectroscopie de corrélation de fluorescence (FCS). Cependant, les aberrations chromatiques des optiques utilisées limitent les performances du montage actuel. Une perspective prometteuse serait de combiner le STED à la microscopie à feuille de lumière (SPIM), plus tolérante des défauts d'achromatisme, et nous montrons les premiers résultats de cette approche. Dans la seconde configuration, les aberrations chromatiques ne sont plus un problème puisqu' une seule longueur d'onde est utilisée pour l'excitation (par absorption à deux photons) et la déplétion. En jouant sur la durée de l'impulsion (et donc la valeur de l'intensité crête), un de ces deux procédés peut être favorisé. La fluorescence est excitée à deux photons par une impulsion femtoseconde, puis est éteinte par émission stimulée à un photon avec une impulsion étirée. Nous avons utilisé une technique résolue en temps (Time-Correlated Single Photon Counting - TCSPC) pour étudier l'efficacité de déplétion du colorant DCM en solution. Les simulations numériques montrent que cette méthode peut être appliquée à la microscopie de super résolution. En fin de cette partie, nous présentons les premières images obtenues avec un microscope développé au laboratoire qui permet l'excitation à deux photons et la déplétion à un photon avec une seule longueur d'onde, ainsi que l' amélioration de la résolution observée. Dans ce travail, nous avons donc mis en place expérimentalement, pour la première fois, deux concepts destinés à simplifier en utilisant deux sources laser originales<br>Stimulated emission depletion (STED) is a well-known super-resolution method. In a STED microscope, a doughnut-shaped beam is superimposed with the excitation beam and keeps the fluorophores in the periphery of the excitation spot in a dark state by stimulated emission, thus effectively improving the spatial resolution in a scanning configuration. This technique requires a complex setup since two laser beams, generally from different sources need to be perfectly aligned. In this work we propose two STED configurations that will simplify the setup and reduce the total cost of such a system. The basic idea in both cases is to use the same laser source for both excitation and stimulated emission depletion. In the first setup we have developed an original two-color source based on a microchip Nd-YAG laser. This microchip laser simultaneously delivers sub-ns pulses at two wavelengths, 355 nm (excitation) and 532 nm (depletion), which are generated by harmonic conversion from an Nd-YAG laser emission and offer the advantage of being intrinsically aligned and synchronized. Further work consisted in determining suitable dyes for this particular source. We have built a microscope setup based on this laser source and obtained images with an improved resolution. The confirmation of the reduction of the excitation volume is showed by Fluorescence Correlation Spectroscopy (FCS) measurements. However, the performance of this system is limited by chromatic aberrations. The combination of Selective Plane Illumination Microscopy (SPIM) with STED is considered. In the second setup the chromatic aberrations are no longer a problem since the same wavelength is used for two photon excitation and one photon depletion. By playing on the duration of the pulse (thus the instantaneous intensity), one of these two processes can be favored. Fluorescence was excited by two photon absorption with a femtosecond pulse, then depleted by one photon stimulated emission with a stretched pulse. We used the Time Correlated Single Photon Counting (TCSPC) method to study the depletion efficiency of DCM dye in solution and numerical simulations show that this method can be applied to super-resolved microscopy. In the end we present the preliminary images obtained with a home-built Two-photon Single wavelength STED microscope and the resolution improvement obtained. Further improvements are to be made to the custom microscope. In this work we have experimentally implemented, for the first time, two concepts meant to simplify the STED setups by using original sources
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Arora, Bhupinder S. "Detection of polysaccharides on a bacterial cell surface using Atomic Force Microscopy." Link to electronic thesis, 2003. http://www.wpi.edu/Pubs/ETD/Available/etd-0826103-011111.
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Thesis (M.S.)--Worcester Polytechnic Institute.<br>Keywords: Leuconostoc mesenteroides NIRC1542; Atomic Force Microscope; Pseudomonas putida KT2442; Adhesion. Includes bibliographical references (p. 75-83).