968 resultados para laser transfection


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Most of the applications of airborne laser scanner data to forestry require that the point cloud be normalized, i.e., each point represents height from the ground instead of elevation. To normalize the point cloud, a digital terrain model (DTM), which is derived from the ground returns in the point cloud, is employed. Unfortunately, extracting accurate DTMs from airborne laser scanner data is a challenging task, especially in tropical forests where the canopy is normally very thick (partially closed), leading to a situation in which only a limited number of laser pulses reach the ground. Therefore, robust algorithms for extracting accurate DTMs in low-ground-point-densitysituations are needed in order to realize the full potential of airborne laser scanner data to forestry. The objective of this thesis is to develop algorithms for processing airborne laser scanner data in order to: (1) extract DTMs in demanding forest conditions (complex terrain and low number of ground points) for applications in forestry; (2) estimate canopy base height (CBH) for forest fire behavior modeling; and (3) assess the robustness of LiDAR-based high-resolution biomass estimation models against different field plot designs. Here, the aim is to find out if field plot data gathered by professional foresters can be combined with field plot data gathered by professionally trained community foresters and used in LiDAR-based high-resolution biomass estimation modeling without affecting prediction performance. The question of interest in this case is whether or not the local forest communities can achieve the level technical proficiency required for accurate forest monitoring. The algorithms for extracting DTMs from LiDAR point clouds presented in this thesis address the challenges of extracting DTMs in low-ground-point situations and in complex terrain while the algorithm for CBH estimation addresses the challenge of variations in the distribution of points in the LiDAR point cloud caused by things like variations in tree species and season of data acquisition. These algorithms are adaptive (with respect to point cloud characteristics) and exhibit a high degree of tolerance to variations in the density and distribution of points in the LiDAR point cloud. Results of comparison with existing DTM extraction algorithms showed that DTM extraction algorithms proposed in this thesis performed better with respect to accuracy of estimating tree heights from airborne laser scanner data. On the other hand, the proposed DTM extraction algorithms, being mostly based on trend surface interpolation, can not retain small artifacts in the terrain (e.g., bumps, small hills and depressions). Therefore, the DTMs generated by these algorithms are only suitable for forestry applications where the primary objective is to estimate tree heights from normalized airborne laser scanner data. On the other hand, the algorithm for estimating CBH proposed in this thesis is based on the idea of moving voxel in which gaps (openings in the canopy) which act as fuel breaks are located and their height is estimated. Test results showed a slight improvement in CBH estimation accuracy over existing CBH estimation methods which are based on height percentiles in the airborne laser scanner data. However, being based on the idea of moving voxel, this algorithm has one main advantage over existing CBH estimation methods in the context of forest fire modeling: it has great potential in providing information about vertical fuel continuity. This information can be used to create vertical fuel continuity maps which can provide more realistic information on the risk of crown fires compared to CBH.

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Hybridihitsaukseksi nimitetään yleensä hitsausprosesseja, joissa yhdistetään kaari- ja laserhitsauksen vahvuudet. Laser- MIG/MAG- prosessit ovat näistä selvästi yleisimpiä, mutta muillekin prosesseille on sovelluksia. Tässä kandidaatintyössä on arvioitu laser- plasma- hybridihitsausprosessin käytettävyyttä teollisuuden prosesseissa. Arviointi on tehty vertailevana kirjallisuustutkimuksena laser- plasma- hybridiprosessin, laserhitsauksen, sekä muiden laser- hybridihitsausprosessien kesken. Prosessien vertailun lisäksi työssä on käsitelty hybridihitsausprosessien parametrien valintaa ja laser- plasma- hybridiprosessin erityispiirteitä.

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There exist several researches and applications about laser welding monitoring and parameter control but not a single one have been created for controlling of laser scribing processes. Laser scribing is considered to be very fast and accurate process and thus it would be necessary to develop accurate turning and monitoring system for such a process. This research focuses on finding out whether it would be possible to develop real-time adaptive control for ultra-fast laser scribing processes utilizing spectrometer online monitoring. The thesis accurately presents how control code for laser parameter tuning is developed using National Instrument's LabVIEW and how spectrometer is being utilized in online monitoring. Results are based on behavior of the control code and accuracy of the spectrometer monitoring when scribing different steel materials. Finally control code success is being evaluated and possible development ideas for future are presented.

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Currently, laser scribing is growing material processing method in the industry. Benefits of laser scribing technology are studied for example for improving an efficiency of solar cells. Due high-quality requirement of the fast scribing process, it is important to monitor the process in real time for detecting possible defects during the process. However, there is a lack of studies of laser scribing real time monitoring. Commonly used monitoring methods developed for other laser processes such a laser welding, are sufficient slow and existed applications cannot be implemented in fast laser scribing monitoring. The aim of this thesis is to find a method for laser scribing monitoring with a high-speed camera and evaluate reliability and performance of the developed monitoring system with experiments. The laser used in experiments is an IPG ytterbium pulsed fiber laser with 20 W maximum average power and Scan head optics used in the laser is Scanlab’s Hurryscan 14 II with an f100 tele-centric lens. The camera was connected to laser scanner using camera adapter to follow the laser process. A powerful fully programmable industrial computer was chosen for executing image processing and analysis. Algorithms for defect analysis, which are based on particle analysis, were developed using LabVIEW system design software. The performance of the algorithms was analyzed by analyzing a non-moving image from the scribing line with resolution 960x20 pixel. As a result, the maximum analysis speed was 560 frames per second. Reliability of the algorithm was evaluated by imaging scribing path with a variable number of defects 2000 mm/s when the laser was turned off and image analysis speed was 430 frames per second. The experiment was successful and as a result, the algorithms detected all defects from the scribing path. The final monitoring experiment was performed during a laser process. However, it was challenging to get active laser illumination work with the laser scanner due physical dimensions of the laser lens and the scanner. For reliable error detection, the illumination system is needed to be replaced.

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The process of depositing thin films by the use of pulsed laser deposition (PLD) has become a more widely used technique for the growth of substances in a thin film form. Pulsed laser deposition allows for the stoichiometric film growth of the target which is of great significance in the deposition of High Temperature Superconducting materials. We will describe a system designed using an excimer laser and vaccum chamber in which thin films and superlattices of YBa2Cuj07_i, PrBa2Cu307_i, and YBajCujOr-j/ PrBajCusOr-^ were deposited on SrTiOs. Results of resistivity measurements using the four probe technique will be shown.

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Gene therapy is predicated upon efficient gene transfer. While viral vectors are the method of choice for transformation efficiency, the immunogenicity and safety concerns remain problematic. Non-viral vectors, on the other hand, have shown high degrees of safety and are mostly non-immunogenic in nature. However, non-viral vectors usually suffer from low levels oftransformation efficiency and transgene expression. Thus, increasing transformation efficiency ofnon-viral vectors, in particular by calcium phosphate co-precipitation technique, is a way of generating a suitable vector for gene therapy and is the aim of this study. It is a long known fact that different cell lines have different transfection efficiencies regardless oftransfection methodology (Lin et a!., 1994). Using commonly available cell lines Madine-Darby Bovine Kidney (MDBK), HeLa and Human Embryonic Kidney (HEK-293), we have shown a decreasing trend ofDNase activity based on a plasmid digestion assay. From densitometry studies, as much as a 40% reduction in DNase activity was observed when comparing HEK-293 (least active) to MDBK (most active). Using various biochemical assays, it was determined that DNase y, in particular, was expressed more highly in MDBK cells than both HeLa and HEK-293. Upon cloning of the bovine DNase y gene, we utilized the sequence information to construct antisense expressing plasmids via both traditional antisense RNA (pASDGneoM) and siRNA (psiRNA-S4, psiRNA-S11 and psiRNA-S16). For the construction ofpASDGneoM, the 3' end of the DNase y was inserted in opposite orientation under a cytomegalovirus (CMV) promoter such that the expression ofRNA complementary to the DNase 2 ymRNA occurred. For siRNA plasmids, the sequence was screened to yield optimal short sequences for siRNA inhibition. The silencing ofbovine DNase y led to an increase in transfection efficiency based on traditional calcium phosphate co-precipitation technique; stable clones of siRNA-producing MDBK cell lines (psiRNA-S4 Bland psiRNA-S4 B4) both demol).strated 4-fold increases in transfection efficiency. Furthermore, serial transfection of antisense DNase y plasmid pASDGneoM and reporter pCMV-~ showed a maximum of 8-fold increase in transfection efficiency when the two separate transfections were carried out 4 hours apart (i.e. transfection ofpASDGneoM, separated by four hours, then transfection ofpCMV-~). Together, these results demonstrate the involvement ofDNase y in reducing transfection efficiency, at least by traditional calcium phosphate technique.

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A Czerny Mount double monochromator is used to measure Raman scattered radiation near 90" from a crystalline, Silicon sample. Incident light is provided by a mixed gas Kr-Ar laser, operating at 5145 A. The double monochromator is calibrated to true wavelength by comparison of Kr and Ar emission Une positions (A) to grating position (A) display [1]. The relationship was found to be hnear and can be described by, y = 1.219873a; - 1209.32, (1) where y is true wavelength (A) and xis grating position display (A). The Raman emission spectra are collected via C"*""*" encoded software, which displays a mV signal from a Photodetector and allows stepping control of the gratings via an A/D interface. [2] The software collection parameters, detector temperature and optics are optimised to yield the best quality spectra. The inclusion of a cryostat allows for temperatmre dependent capabihty ranging from 4 K to w 350 K. Silicon Stokes temperatm-e dependent Raman spectra, generally show agreement with Uterature results [3] in their frequency haxdening, FWHM reduction and intensity increase as temperature is reduced. Tests reveal that a re-alignment of the double monochromator is necessary before spectral resolution can approach literature standard. This has not yet been carried out due to time constraints.

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Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency.

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The regenerating amphibian limb provides a useful system for studying genes involved in the establishment of positional information. While a number of candidate genes that may playa role in pattern formation have been identified, their function in vivo is unknown in this system. To better ascertain the role of these genes, it would be useful to be able to alter their normal patterns of expression in vivo and to assess the effects of this misexpression on limb pattern. In order to achieve this, a method of introducing a plasmid containing the eDNA of a gene of interest into a newt blastema (a growth zone of mesenchymal progenitor cells) is needed. Unfortunately, most commonly used transfection techniques cannot be used with newt blastema cells. In this study, I have used the techniques of lipofection and direct gene transfer to introduce plasmid DNA containing reporter genes into the cells of a regenerating newt limb. The technique of lipofection was most effective when the blastema cells were transfected in vitro. The optimal ratio for transfection was shown to be 1:3 DNA:Lipofectin (W/w) , and an increase in the amount of DNA present in the mixture (1:3 ratio maintained) resulted in a corresponding increase in gene expression. The technique of direct gene transfer was used to transfect newt blastema cells with and without prior complex formation with Lipofectin. Injection of plasmid DNA alone provided the most 3 promising results. It was possible to introduce plasmid DNA containing the reporter gene ~-galactosidase and achieve significant gene expression in cells associated with the injection site. In the future, it would be interesting to use this technique to inject plasmid DNA containing a gene which may have a role in pattern formation into specific areas of the newt blastema and to analyze the resulting limb pattern that emerges.

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SrMg^Rui-iOa thin films were made by using pulsed laser deposition on SrTiOa (100) substrates in either O2 or Ar atmosphere. The thin films were characterized by x-ray diffraction, energy dispersive x-ray microanalysis, dc resistivity measurement, and dc magnetization measurement. The effect of Mg doping was observed. As soon as the amount of Mg increased in SrMg-cRui-iOa thin films, the magnetization decreased, and the resistivity increased. It had little effect on the Curie temperature (transition temperature). The magnetization states of SrMgiRui-iOa thin films, for x < 0.15, are similar to SrRuOs films. X-ray diffraction results for SrMga-Rui-iOa thin films made in oxygen showed that the films are epitaxial. The thin films could not be well made in Ar atmosphere during laser ablation as there was no clear peak of SrMg^Rui-iOa in x-ray diffraction results. Substrate temperatures had an effect on the resistivity of the films. The residual resistivity ratios were increased by increasing substrate temperature. It was observed that the thickness of thin films are another factor for film quality: Thin films were epitaxial, but thicker films were not epitaxial.

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Tesis (Maestría en Ciencias en Ingeniería Eléctrica con Especialidad en Potencia) UANL

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La caractérisation de matériaux par spectroscopie optique d’émission d’un plasma induit par laser (LIPS) suscite un intérêt qui ne va que s’amplifiant, et dont les applications se multiplient. L’objectif de ce mémoire est de vérifier l’influence du choix des raies spectrales sur certaines mesures du plasma, soit la densité électronique et la température d’excitation des atomes neutres et ionisés une fois, ainsi que la température d’ionisation. Nos mesures sont intégrées spatialement et résolues temporellement, ce qui est typique des conditions opératoires du LIPS, et nous avons utilisé pour nos travaux des cibles binaires d’aluminium contenant des éléments à l’état de trace (Al-Fe et Al-Mg). Premièrement, nous avons mesuré la densité électronique à l’aide de l’élargissement Stark de raies de plusieurs espèces (Al II, Fe II, Mg II, Fe I, Mg I, Halpha). Nous avons observé que les densités absolues avaient un comportement temporel différent en fonction de l’espèce. Les raies ioniques donnent des densités électroniques systématiquement plus élevées (jusqu’à 50 % à 200 ns après l’allumage du plasma), et décroissent plus rapidement que les densités issues des raies neutres. Par ailleurs, les densités obtenues par les éléments traces Fe et Mg sont moindres que les densités obtenues par l’observation de la raie communément utilisée Al II à 281,618 nm. Nous avons parallèlement étudié la densité électronique déterminée à l’aide de la raie de l’hydrogène Halpha, et la densité électronique ainsi obtenue a un comportement temporel similaire à celle obtenue par la raie Al II à 281,618 nm. Les deux espèces partagent probablement la même distribution spatiale à l’intérieur du plasma. Finalement, nous avons mesuré la température d’excitation du fer (neutre et ionisé, à l’état de trace dans nos cibles), ainsi que la température d’ionisation, à l’aide de diagrammes de Boltzmann et de Saha-Boltzmann, respectivement. À l’instar de travaux antérieurs (Barthélémy et al., 2005), il nous est apparu que les différentes températures convergeaient vers une température unique (considérant nos incertitudes) après 2-3 microsecondes. Les différentes températures mesurées de 0 à 2 microsecondes ne se recoupent pas, ce qui pourrait s’expliquer soit par un écart à l’équilibre thermodynamique local, soit en considérant un plasma inhomogène où la distribution des éléments dans la plume n’est pas similaire d’un élément à l’autre, les espèces énergétiques se retrouvant au cœur du plasma, plus chaud, alors que les espèces de moindre énergie se retrouvant principalement en périphérie.

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La technique du laser femtoseconde (fs) a été précédemment utilisée pour la production de nanoparticules d'or dans un environnement aqueux biologiquement compatible. Au cours de ce travail de maîtrise, cette méthode a été investiguée en vue d'une application pour la fabrication de nanocristaux de médicament en utilisant le paclitaxel comme modèle. Deux procédés distincts de cette technologie à savoir l'ablation et la fragmentation ont été étudiés. L'influence de la puissance du laser, de point de focalisation, et de la durée du traitement sur la distribution de taille des particules obtenues ainsi que leur intégrité chimique a été évaluée. Les paramètres ont ainsi été optimisés pour la fabrication des nanoparticules. L’évaluation morphologique et chimique a été réalisée par microscopie électronique et spectroscopie infrarouge respectivement. L'état cristallin des nanoparticules de paclitaxel a été caractérisé par calorimétrie differentielle et diffraction des rayons X. L'optimisation du procédé de production de nanoparticules par laser fs a permis d'obtenir des nanocristaux de taille moyenne (400 nm, polydispersité ≤ 0,3). Cependant une dégradation non négligeable a été observée. La cristallinité du médicament a été maintenue durant la procédure de réduction de taille, mais le paclitaxel anhydre a été transformé en une forme hydratée. Les résultats de cette étude suggèrent que le laser fs peut générer des nanocristaux de principe actif. Cependant cette technique peut se révéler problématique pour des médicaments sensibles à la dégradation. Grâce à sa facilité d'utilisation et la possibilité de travailler avec des quantités restreintes de produit, le laser fs pourrait représenter une alternative valable pour la production de nanoparticules de médicaments peu solubles lors des phases initiales de développement préclinique. Mots-clés: paclitaxel, nanocristaux, laser femtoseconde, ablation, fragmentation

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La débitmétrie au laser à effet Doppler (LDF) constitue une méthode prometteuse et non-invasive pour l'étude du débit sanguin local dans l'œil. Cette technique est basée sur un changement de fréquence subi par la lumière lors du mouvement des globules rouges dans les vaisseaux. Une nouvelle sonde LDF a été testée pour sa sensibilité à évaluer la circulation rétinienne/choroïdienne sous des conditions hypercapniques et en présence de diverses substances vasoactives ou suivant la photocoagulation des artères rétiniennes chez le rat. Après dilatation pupillaire, la sonde LDF a été placée en contact avec la cornée de rats anesthésiés et parallèle à l'axe optique. L'hypercapnie a été provoquée par inhalation de CO2 (8% dans de l'air médical), alors que les agents pharmacologiques ont été injectés de façon intravitréenne. La contribution relative à la circulation choroïdienne a été évaluée à la suite de la photocoagulation des artères rétiniennes. Le débit sanguin s'est trouvé significativement augmenté à la suite de l'hypercanie (19%), de l'adénosine (14%) ou du nitroprusside de sodium (16%) comparativement au niveau de base, alors que l'endothéline-1 a provoqué une baisse du débit sanguin (11%). La photocoagulation des artères rétiniennes a significativement diminué le débit sanguin (33%). Des mesures en conditions pathologiques ont ensuite été obtenues après l'injection intravitréenne d'un agoniste sélectif du récepteur B1 (RB1). Ce récepteur des kinines est surexprimé dans la rétine des rats rendus diabétiques avec la streptozotocine (STZ) en réponse à l'hyperglycémie et au stress oxydatif. Les résultats ont montré que le RB1 est surexprimé dans la rétine chez les rats diabétiques-STZ à 4 jours et 6 semaines. À ces moments, le débit sanguin rétinien/choroïdien a été significativement augmenté (15 et 18 %) après l'injection de l'agoniste, suggérant un effet vasodilatateur des RB1 dans l'œil diabétique. Bien que la circulation choroïdienne contribue probablement au signal LDF, les résultats démontrent que le LDF représente une technique efficace et non-invasive pour l'étude de la microcirculation rétinienne in-vivo en continu. Cette méthode peut donc être utilisée pour évaluer de façon répétée les réponses du débit sanguin pendant des modifications métaboliques ou pharmacologiques dans des modèles animaux de maladies oculaires.

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L’ablation de cibles d’Al nanocristallines (taille moyenne des cristallites d = 3,1 et 6,2 nm) par impulsions laser ultrabrèves (200 fs) a été étudiée par l’entremise de si- mulations combinant la dynamique moléculaire et le modèle à deux températures (two- temperature model, TTM) pour des fluences absorbées allant de 100 à 1300 J/m2. Nos simulations emploient un potentiel d’interaction de type EAM et les propriétés électro- niques des cibles en lien avec le TTM sont représentées par un modèle réaliste possédant une forme distincte dans le solide monocristallin, le solide nanocristallin et le liquide. Nous avons considéré l’effet de la taille moyenne des cristallites de même que celui de la porosité et nous avons procédé à une comparaison directe avec des cibles mono- cristallines. Nous avons pu montrer que le seuil d’ablation des métaux nanocristallins est significativement plus bas, se situant à 400 J/m2 plutôt qu’à 600 J/m2 dans le cas des cibles monocristallines, l’écart étant principalement dû à l’onde mécanique plus im- portante présente lors de l’ablation. Leur seuil de spallation de la face arrière est aussi significativement plus bas de par la résistance à la tension plus faible (5,40 GPa contre 7,24 GPa) des cibles nanocristallines. Il est aussi apparu que les contraintes résiduelles accompagnant généralement l’ablation laser sont absentes lors de l’ablation de cibles d’aluminium nanocristallines puisque la croissance cristalline leur permet d’abaisser leur volume spécifique. Nos résultats indiquent aussi que le seuil de fusion des cibles nano- cristallines est réduit de façon marquée dans ces cibles ce qui s’explique par la plus faible énergie de cohésion inhérente à ces matériaux. Nos simulations permettent de montrer que les propriétés structurelles et électroniques propres aux métaux nanocristallins ont toutes deux un impact important sur l’ablation.