Detecção de regiões de via integrando dados de varredura a LASER aerotransportado e imagens de alta resolução

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Reconstrução de contornos e cumeeiras de telhados de edifícios a partir de imagens aéreas de alta-resolução e poliedros representativos de edifícios extraídos de dados LASER

Pós-graduação em Ciências Cartográficas - FCT

Refinamento geométrico de contornos e cumeeiras de telhados de edifícios extraídos de dados laser com uso de imagem aérea

In this paper, a method is proposed to refine the LASER 3D roofs geometrically by using a high-resolution aerial image and Markov Random Field (MRF) models. In order to do so, a MRF description for grouping straight lines is developed, assuming that each projected side contour and ridge is topologically correct and that it is only necessary to improve its accuracy. Although the combination of laser data with data from image is most justified for refining roof contour, the structure of ridges can give greater robustness in the topological description of the roof structure. The MRF model is formulated based on relationships (length, proximity, and orientation) between the straight lines extracted from the image and projected polygon and also on retangularity and corner injunctions. The energy function associated with MRF is minimized by the genetic algorithm optimization method, resulting in the grouping of straight lines for each roof object. Finally, each grouping of straight lines is topologically reconstructed based on the topology of the corresponding LASER scanning polygon projected onto the image-space. The results obtained were satisfactory. This method was able to provide polygons roof refined buildings in which most of its contour sides and ridges were geometrically improved.

Monitoramento e avaliação de processos erosivos marginais em reservatórios de usinas hidrelétricas por meio de varredura a laser

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Montagem e avaliação de um sistema de varredura a LASER embarcado em VANT

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Montagem e avaliação de um sistema de varredura a LASER embarcado em VANT

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Microtensile bond strength of resin composite to dentin treated with Er:YAG laser of bleached teeth

The objective of this study was to evaluate the influence of Er:YAG laser (lambda = 2.94 mu m) on microtensile bond strength (mu TBS) and superficial morphology of bovine dentin bleached with 16% carbamide peroxide. Forty bovine teeth blocks (7 x 3 x 3 mm(3)) were randomly assigned to four groups: G1- bleaching and Er:YAG irradiation with energy density of 25.56 J/cm(2) (focused mode); G2 - bleaching; G3 - no-bleaching and Er:YAG irradiation (25.56 J/cm(2)); G4 - control, non-treated. G1 and G2 were bleached with 16% carbamide peroxide for 6 h during 21 days. Afterwards, all blocks were abraded with 320 to 600-grit abrasive papers to obtain flat standardized dentin surfaces. G1 and G3 were Er:YAG irradiated. Blocks were immediately restored with 4-mm-high composite resin (Adper Single Bond 2, Z-250-3 M/ESPE). After 24 h, the restored blocks (n = 9) were serially sectioned and trimmed to an hour-glass shape of approximately 1 mm(2) at the bonded interface area, and tested in tension in a universal testing machine (1 mm/ min). Failure mode was determined at a magnification of 100x using a stereomicroscope. One block of each group was selected for scanning electron microscope (SEM) analysis. mu TBS data was analyzed by two-way ANOVA and Tukey test (alpha = 0.05). Mean bond strengths (SD) in MPa were: G1- 32.7 (5.9)(A); G2- 31.1 (6.3)(A); G3- 25.2 (8.3)(B); G4- 36.7 (9.9).(A) Groups with different uppercase letters were significantly different from each other (p < .05). Enamel bleaching procedure did not affect mu TBS values for dentin adhesion. Er:YAG laser irradiation with 25.56 J/cm(2) prior to adhesive procedure of bleached teeth did not affect mu TBS at dentin and promoted a dentin surface with no smear layer and opened dentin tubules observed under SEM. On the other hand, Er:YAG laser irradiation prior to adhesive procedure of non-bleached surface impaired mu TBS compared to the control group.

Analyse der intrazellulären Freisetzung von Wirkstoffmodellen via konfokaler Laser-Raster-Mikroskopie

In der vorliegenden Arbeit wurde gezeigt, wie man das Potential nanopartikulärer Systeme, die vorwiegend via Miniemulsion hergestellt wurden, im Hinblick auf „Drug Delivery“ ausnutzen könnte, indem ein Wirkstoffmodell auf unterschiedliche Art und Weise intrazellulär freigesetzt wurde. Dies wurde hauptsächlich mittels konfokaler Laser-Raster-Mikrokopie (CLSM) in Kombination mit dem Bildbearbeitungsprogramm Volocity® analysiert.rnPBCA-Nanokapseln eigneten sich besonders, um hydrophile Substanzen wie etwa Oligonukleotide zu verkapseln und sie so auf ihrem Transportweg in die Zellen vor einem etwaigen Abbau zu schützen. Es konnte eine Freisetzung der Oligonukleotide in den Zellen aufgrund der elektrostatischen Anziehung des mitochondrialen Membranpotentials nachgewiesen werden. Dabei war die Kombination aus Oligonukleotid und angebundenem Cyanin-Farbstoff (Cy5) an der 5‘-Position der Oligonukleotid-Sequenz ausschlaggebend. Durch quantitative Analysen mittels Volocity® konnte die vollständige Kolokalisation der freigesetzten Oligonukleotide an Mitochondrien bewiesen werden, was anhand der Kolokalisationskoeffizienten „Manders‘ Coefficients“ M1 und M2 diskutiert wurde. Es konnte ebenfalls aufgrund von FRET-Studien doppelt markierter Oligos gezeigt werden, dass die Oligonukleotide weder beim Transport noch bei der Freisetzung abgebaut wurden. Außerdem wurde aufgeklärt, dass nur der Inhalt der Nanokapseln, d. h. die Oligonukleotide, an Mitochondrien akkumulierte, das Kapselmaterial selbst jedoch in anderen intrazellulären Bereichen aufzufinden war. Eine Kombination aus Cyanin-Farbstoffen wie Cy5 mit einer Nukleotidsequenz oder einem Wirkstoff könnte also die Basis für einen gezielten Wirkstofftransport zu Mitochondrien liefern bzw. die Grundlage schaffen, eine Freisetzung aus Kapseln ins Zytoplasma zu gewährleisten.rnDer vielseitige Einsatz der Miniemulsion gestattete es, nicht nur Kapseln sondern auch Nanopartikel herzustellen, in welchen hydrophobe Substanzen im Partikelkern eingeschlossen werden konnten. Diese auf hydrophobe Wechselwirkungen beruhende „Verkapselung“ eines Wirkstoffmodells, in diesem Fall PMI, wurde bei PDLLA- bzw. PS-Nanopartikeln ausgenutzt, welche durch ein HPMA-basiertes Block-Copolymer stabilisiert wurden. Dabei konnte gezeigt werden, dass das hydrophobe Wirkstoffmodell PMI innerhalb kürzester Zeit in die Zellen freigesetzt wurde und sich in sogenannte „Lipid Droplets“ einlagerte, ohne dass die Nanopartikel selbst aufgenommen werden mussten. Daneben war ein intrazelluläres Ablösen des stabilisierenden Block-Copolymers zu verzeichnen, welches rn8 h nach Partikelaufnahme erfolgte und ebenfalls durch Analysen mittels Volocity® untermauert wurde. Dies hatte jedoch keinen Einfluss auf die eigentliche Partikelaufnahme oder die Freisetzung des Wirkstoffmodells. Ein großer Vorteil in der Verwendung des HPMA-basierten Block-Copolymers liegt darin begründet, dass auf zeitaufwendige Waschschritte wie etwa Dialyse nach der Partikelherstellung verzichtet werden konnte, da P(HPMA) ein biokompatibles Polymer ist. Auf der anderen Seite hat man aufgrund der Syntheseroute dieses Block-Copolymers vielfältige Möglichkeiten, Funktionalitäten wie etwa Fluoreszenzmarker einzubringen. Eine kovalente Anbindung eines Wirkstoffs ist ebenfalls denkbar, welcher intrazellulär z. B. aufgrund von enzymatischen Abbauprozessen langsam freigesetzt werden könnte. Somit bietet sich die Möglichkeit mit Nanopartikeln, die durch HPMA-basierte Block-Copolymere stabilisiert wurden, gleichzeitig zwei unterschiedliche Wirkstoffe in die Zellen zu bringen, wobei der eine schnell und der zweite über einen längeren Zeitraum hinweg (kontrolliert) freigesetzt werden könnte.rnNeben Nanokapseln sowie –partikeln, die durch inverse bzw. direkte Miniemulsion dargestellt wurden, sind auch Nanohydrogelpartikel untersucht worden, die sich aufgrund von Selbstorganisation eines amphiphilen Bock-Copolymers bildeten. Diese Nanohydrogelpartikel dienten der Komplexierung von siRNA und wurden hinsichtlich ihrer Anreicherung in Lysosomen untersucht. Aufgrund der Knockdown-Studien von Lutz Nuhn konnte ein Unterschied in der Knockdown-Effizienz festgestellt werden, je nach dem, ob 100 nm oder 40 nm große Nanohydrogelpartikel verwendet wurden. Es sollte festgestellt werden, ob eine größenbedingte, unterschiedlich schnelle Anreicherung dieser beiden Partikel in Lysosomen erfolgte, was die unterschiedliche Knockdown-Effizienz erklären könnte. CLSM-Studien und quantitative Kolokalisationsstudien gaben einen ersten Hinweis auf diese Größenabhängigkeit. rnBei allen verwendeten nanopartikulären Systemen konnte eine Freisetzung ihres Inhalts gezeigt werden. Somit bieten sie ein großes Potential als Wirkstoffträger für biomedizinische Anwendungen.rn

Experimental and numerical study of cw green laser crystallization of a-Si:H thin films

Crystallization and grain growth technique of thin film silicon are among the most promising methods for improving efficiency and lowering cost of solar cells. A major advantage of laser crystallization and annealing over conventional heating methods is its ability to limit rapid heating and cooling to thin surface layers. Laser energy is used to heat the amorphous silicon thin film, melting it and changing the microstructure to polycrystalline silicon (poly-Si) as it cools. Depending on the laser density, the vaporization temperature can be reached at the center of the irradiated area. In these cases ablation effects are expected and the annealing process becomes ineffective. The heating process in the a-Si thin film is governed by the general heat transfer equation. The two dimensional non-linear heat transfer equation with a moving heat source is solve numerically using the finite element method (FEM), particularly COMSOL Multiphysics. The numerical model help to establish the density and the process speed range needed to assure the melting and crystallization without damage or ablation of the silicon surface. The samples of a-Si obtained by physical vapour deposition were irradiated with a cw-green laser source (Millennia Prime from Newport-Spectra) that delivers up to 15 W of average power. The morphology of the irradiated area was characterized by confocal laser scanning microscopy (Leica DCM3D) and Scanning Electron Microscopy (SEM Hitachi 3000N). The structural properties were studied by micro-Raman spectroscopy (Renishaw, inVia Raman microscope).

Auscultación de presas de hormigón con láser escáner terrestre y modelización de las deformaciones del paramento

Se está produciendo en la geodesia un cambio de paradigma en la concepción de los modelos digitales del terreno, pasando de diseñar el modelo con el menor número de puntos posibles a hacerlo con cientos de miles o millones de puntos. Este cambio ha sido consecuencia de la introducción de nuevas tecnologías como el escáner láser, la interferometría radar y el tratamiento de imágenes. La rápida aceptación de estas nuevas tecnologías se debe principalmente a la gran velocidad en la toma de datos, a la accesibilidad por no precisar de prisma y al alto grado de detalle de los modelos. Los métodos topográficos clásicos se basan en medidas discretas de puntos que considerados en su conjunto forman un modelo; su precisión se deriva de la precisión en la toma singular de estos puntos. La tecnología láser escáner terrestre (TLS) supone una aproximación diferente para la generación del modelo del objeto observado. Las nubes de puntos, producto del escaneo con TLS, pasan a ser tratadas en su conjunto mediante análisis de áreas, de forma que ahora el modelo final no es el resultado de una agregación de puntos sino la de la mejor superficie que se adapta a las nubes de puntos. Al comparar precisiones en la captura de puntos singulares realizados con métodos taquimétricos y equipos TLS la inferioridad de estos últimos es clara; sin embargo es en el tratamiento de las nubes de puntos, con los métodos de análisis basados en áreas, se han obtenido precisiones aceptables y se ha podido considerar plenamente la incorporación de esta tecnología en estudios de deformaciones y movimientos de estructuras. Entre las aplicaciones del TLS destacan las de registro del patrimonio, registro de las fases en la construcción de plantas industriales y estructuras, atestados de accidentes y monitorización de movimientos del terreno y deformaciones de estructuras. En la auscultación de presas, comparado con la monitorización de puntos concretos dentro, en coronación o en el paramento de la presa, disponer de un modelo continuo del paramento aguas abajo de la presa abre la posibilidad de introducir los métodos de análisis de deformaciones de superficies y la creación de modelos de comportamiento que mejoren la comprensión y previsión de sus movimientos. No obstante, la aplicación de la tecnología TLS en la auscultación de presas debe considerarse como un método complementario a los existentes. Mientras que los péndulos y la reciente técnica basada en el sistema de posicionamiento global diferencial (DGPS) dan una información continua de los movimientos de determinados puntos de la presa, el TLS permite ver la evolución estacional y detectar posibles zonas problemáticas en todo el paramento. En este trabajo se analizan las características de la tecnología TLS y los parámetros que intervienen en la precisión final de los escaneos. Se constata la necesidad de utilizar equipos basados en la medida directa del tiempo de vuelo, también llamados pulsados, para distancias entre 100 m y 300 m Se estudia la aplicación del TLS a la modelización de estructuras y paramentos verticales. Se analizan los factores que influyen en la precisión final, como el registro de nubes, tipo de dianas y el efecto conjunto del ángulo y la distancia de escaneo. Finalmente, se hace una comparación de los movimientos dados por los péndulos directos de una presa con los obtenidos del análisis de las nubes de puntos correspondientes a varias campañas de escaneos de la misma presa. Se propone y valida el empleo de gráficos patrón para relacionar las variables precisión o exactitud con los factores distancia y ángulo de escaneo en el diseño de trabajos de campo. Se expone su aplicación en la preparación del trabajo de campo para la realización de una campaña de escaneos dirigida al control de movimientos de una presa y se realizan recomendaciones para la aplicación de la técnica TLS a grandes estructuras. Se ha elaborado el gráfico patrón de un equipo TLS concreto de alcance medio. Para ello se hicieron dos ensayos de campo en condiciones reales de trabajo, realizando escaneos en todo el rango de distancias y ángulos de escaneo del equipo. Se analizan dos métodos para obtener la precisión en la modelización de paramentos y la detección de movimientos de estos: el método del “plano de mejor ajuste” y el método de la “deformación simulada”. Por último, se presentan los resultados de la comparación de los movimientos estacionales de una presa arco-gravedad entre los registrados con los péndulos directos y los obtenidos a partir de los escaneos realizados con un TLS. Los resultados muestran diferencias de milímetros, siendo el mejor de ellos del orden de un milímetro. Se explica la metodología utilizada y se hacen consideraciones respecto a la densidad de puntos de las nubes y al tamaño de las mallas de triángulos. A shift of paradigm in the conception of the survey digital models is taking place in geodesy, moving from designing a model with the fewer possible number of points to models of hundreds of thousand or million points. This change has happened because of the introduction of new technologies like the laser scanner, the interferometry radar and the processing of images. The fast acceptance of these new technologies has been due mainly to the great speed getting the data, to the accessibility as reflectorless technique, and to the high degree of detail of the models. Classic survey methods are based on discreet measures of points that, considered them as a whole, form a model; the precision of the model is then derived from the precision measuring the single points. The terrestrial laser scanner (TLS) technology supposes a different approach to the model generation of the observed object. Point cloud, the result of a TLS scan, must be treated as a whole, by means of area-based analysis; so, the final model is not an aggregation of points but the one resulting from the best surface that fits with the point cloud. Comparing precisions between the one resulting from the capture of singular points made with tachometric measurement methods and with TLS equipment, the inferiority of this last one is clear; but it is in the treatment of the point clouds, using area-based analysis methods, when acceptable precisions have been obtained and it has been possible to consider the incorporation of this technology for monitoring structures deformations. Among TLS applications it have to be emphasized those of registry of the cultural heritage, stages registry during construction of industrial plants and structures, police statement of accidents and monitorization of land movements and structures deformations. Compared with the classical dam monitoring, approach based on the registry of a set of points, the fact having a continuous model of the downstream face allows the possibility of introducing deformation analysis methods and behavior models that would improve the understanding and forecast of dam movements. However, the application of TLS technology for dam monitoring must be considered like a complementary method with the existing ones. Pendulums and recently the differential global positioning system (DGPS) give a continuous information of the movements of certain points of the dam, whereas TLS allows following its seasonal evolution and to detect damaged zones of the dam. A review of the TLS technology characteristics and the factors affecting the final precision of the scanning data is done. It is stated the need of selecting TLS based on the direct time of flight method, also called pulsed, for scanning distances between 100m and 300m. Modelling of structures and vertical walls is studied. Factors that influence in the final precision, like the registry of point clouds, target types, and the combined effect of scanning distance and angle of incidence are analyzed. Finally, a comparison among the movements given by the direct pendulums of a dam and the ones obtained from the analysis of point clouds is done. A new approach to obtain a complete map-type plot of the precisions of TLS equipment based on the direct measurement of time of flight method at midrange distances is presented. Test were developed in field-like conditions, similar to dam monitoring and other civil engineering works. Taking advantage of graphic semiological techniques, a “distance - angle of incidence” map based was designed and evaluated for field-like conditions. A map-type plot was designed combining isolines with sized and grey scale points, proportional to the precision values they represent. Precisions under different field conditions were compared with specifications. For this purpose, point clouds were evaluated under two approaches: the standar "plane-of-best-fit" and the proposed "simulated deformation”, that showed improved performance. These results lead to a discussion and recommendations about optimal TLS operation in civil engineering works. Finally, results of the comparison of seasonal movements of an arc-gravity dam between the registered by the direct pendulums ant the obtained from the TLS scans, are shown. The results show differences of millimeters, being the best around one millimeter. The used methodology is explained and considerations with respect to the point cloud density and to the size of triangular meshes are done.

Influence of specimens' design and manufacturing process on microtensile bond strength to enamel: laboratory and FEA comparison

This study evaluated the effect of specimens' design and manufacturing process on microtensile bond strength, internal stress distributions (Finite Element Analysis - FEA) and specimens' integrity by means of Scanning Electron Microscopy (SEM) and Laser Scanning Confocal Microscopy (LCM). Excite was applied to flat enamel surface and a resin composite build-ups were made incrementally with 1-mm increments of Tetric Ceram. Teeth were cut using a diamond disc or a diamond wire, obtaining 0.8 mm² stick-shaped specimens, or were shaped with a Micro Specimen Former, obtaining dumbbell-shaped specimens (n = 10). Samples were randomly selected for SEM and LCM analysis. Remaining samples underwent microtensile test, and results were analyzed with ANOVA and Tukey test. FEA dumbbell-shaped model resulted in a more homogeneous stress distribution. Nonetheless, they failed under lower bond strengths (21.83 ± 5.44 MPa)c than stick-shaped specimens (sectioned with wire: 42.93 ± 4.77 MPaª; sectioned with disc: 36.62 ± 3.63 MPa b), due to geometric irregularities related to manufacturing process, as noted in microscopic analyzes. It could be concluded that stick-shaped, nontrimmed specimens, sectioned with diamond wire, are preferred for enamel specimens as they can be prepared in a less destructive, easier, and more precise way.

Sudan Black B treatment reduces autofluorescence and improves resolution of in situ hybridization specific fluorescent signals of brain sections

Interference by autofluorescence is one of the major concerns of immunofluorescence analysis of in situ hybridization-based diagnostic assays. We present a useful technique that reduces autofluorescent background without affecting the tissue integrity or direct immunofluorescence signals in brain sections. Using six different protocols, such as ammonia/ethanol, Sudan Black B (SBB) in 70% ethanol, photobleaching with UV light and different combinations of them in both formalin-fixed paraffin-embedded and frozen human brain tissue sections, we have found that tissue treatment of SBB in a concentration of 0.1% in 70% ethanol is the best approach to reduce/eliminate tissue autofluorescence and background, while preserving the specific fluorescence hybridization signals. This strategy is a feasible, non-time consuming method that provides a reasonable compromise between total reduction of the tissue autofluorescence and maintenance of specific fluorescent labels.

Chrysotile effects on human lung cell carcinoma in culture: 3-D reconstruction and DNA quantification by image analysis

Background: Chrysotile is considered less harmful to human health than other types of asbestos fibers. Its clearance from the lung is faster and, in comparison to amphibole forms of asbestos, chrysotile asbestos fail to accumulate in the lung tissue due to a mechanism involving fibers fragmentation in short pieces. Short exposure to chrysotile has not been associated with any histopathological alteration of lung tissue. Methods: The present work focuses on the association of small chrysotile fibers with interphasic and mitotic human lung cancer cells in culture, using for analyses confocal laser scanning microscopy and 3D reconstructions. The main goal was to perform the analysis of abnormalities in mitosis of fibers-containing cells as well as to quantify nuclear DNA content of treated cells during their recovery in fiber-free culture medium. Results: HK2 cells treated with chrysotile for 48 h and recovered in additional periods of 24, 48 and 72 h in normal medium showed increased frequency of multinucleated and apoptotic cells. DNA ploidy of the cells submitted to the same chrysotile treatment schedules showed enhanced aneuploidy values. The results were consistent with the high frequency of multipolar spindles observed and with the presence of fibers in the intercellular bridge during cytokinesis. Conclusion: The present data show that 48 h chrysotile exposure can cause centrosome amplification, apoptosis and aneuploid cell formation even when long periods of recovery were provided. Internalized fibers seem to interact with the chromatin during mitosis, and they could also interfere in cytokinesis, leading to cytokinesis failure which forms aneuploid or multinucleated cells with centrosome amplification.

The Fate of Chrysotile-Induced Multipolar Mitosis and Aneuploid Population in Cultured Lung Cancer Cells

Chrysotile is one of the six types of asbestos, and it is the only one that can still be commercialized in many countries. Exposure to other types of asbestos has been associated with serious diseases, such as lung carcinomas and pleural mesotheliomas. The association of chrysotile exposure with disease is controversial. However, in vitro studies show the mutagenic potential of chrysotile, which can induce DNA and cell damage. The present work aimed to analyze alterations in lung small cell carcinoma cultures after 48 h of chrysotile exposure, followed by 2, 4 and 8 days of recovery in fiber-free culture medium. Some alterations, such as aneuploid cell formation, increased number of cells in G2/M phase and cells in multipolar mitosis were observed even after 8 days of recovery. The presence of chrysotile fibers in the cell cultures was detected and cell morphology was observed by laser scanning confocal microscopy. After 4 and 8 days of recovery, only a few chrysotile fragments were present in some cells, and the cellular morphology was similar to that of control cells. Cells transfected with the GFP-tagged alpha-tubulin plasmid were treated with chrysotile for 24 or 48 h and cells in multipolar mitosis were observed by time-lapse microscopy. Fates of these cells were established: retention in metaphase, cell death, progression through M phase generating more than two daughter cells or cell fusion during telophase or cytokinesis. Some of them were related to the formation of aneuploid cells and cells with abnormal number of centrosomes.

Invasion of Ureaplasma diversum in Hep-2 cells

Background: Understanding mollicutes is challenging due to their variety and relationship with host cells. Invasion has explained issues related to their opportunistic role. Few studies have been done on the Ureaplasma diversum mollicute, which is detected in healthy or diseased bovine. The invasion in Hep-2 cells of four clinical isolates and two reference strains of their ureaplasma was studied by Confocal Laser Scanning Microscopy and gentamicin invasion assay. Results: The isolates and strains used were detected inside the cells after infection of one minute without difference in the arrangement for adhesion and invasion. The adhesion was scattered throughout the cells, and after three hours, the invasion of the ureaplasmas surrounded the nuclear region but were not observed inside the nuclei. The gentamicin invasion assay detected that 1% of the ATCC strains were inside the infected Hep-2 cells in contrast to 10% to the clinical isolates. A high level of phospholipase C activity was also detected in all studied ureaplasma. Conclusions: The results presented herein will help better understand U. diversum infections, aswell as cellular attachment and virulence.