962 resultados para laser interferometry-based guidance


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Understanding and controlling the mechanism of the diffusion of small molecules, macromolecules and nanoparticles in heterogeneous environments is of paramount fundamental and technological importance. The aim of the thesis is to show, how by studying the tracer diffusion in complex systems, one can obtain information about the tracer itself, and the system where the tracer is diffusing. rnIn the first part of my thesis I will introduce the Fluorescence Correlation Spectroscopy (FCS) which is a powerful tool to investigate the diffusion of fluorescent species in various environments. By using the main advantage of FCS namely the very small probing volume (<1µm3) I was able to track the kinetics of phase separation in polymer blends at late stages by looking on the molecular tracer diffusion in individual domains of the heterogeneous structure of the blend. The phase separation process at intermediate stages was monitored with laser scanning confocal microscopy (LSCM) in real time providing images of droplet coalescence and growth. rnIn a further project described in my thesis I will show that even when the length scale of the heterogeneities becomes smaller than the FCS probing volume one can still obtain important microscopic information by studying small tracer diffusion. To do so, I will introduce a system of star shaped polymer solutions and will demonstrate that the mobility of small molecular tracers on microscopic level is nearly not affected by the transition of the polymer system to a “glassy” macroscopic state. rnIn the last part of the thesis I will introduce and describe a new stimuli responsive system which I have developed, that combines two levels of nanoporosity. The system is based on poly-N-isopropylacrylamide (PNIPAM) and silica inverse opals (iOpals), and allows controlling the diffusion of tracer molecules. rn

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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

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Seventeen bones (sixteen cadaveric bones and one plastic bone) were used to validate a method for reconstructing a surface model of the proximal femur from 2D X-ray radiographs and a statistical shape model that was constructed from thirty training surface models. Unlike previously introduced validation studies, where surface-based distance errors were used to evaluate the reconstruction accuracy, here we propose to use errors measured based on clinically relevant morphometric parameters. For this purpose, a program was developed to robustly extract those morphometric parameters from the thirty training surface models (training population), from the seventeen surface models reconstructed from X-ray radiographs, and from the seventeen ground truth surface models obtained either by a CT-scan reconstruction method or by a laser-scan reconstruction method. A statistical analysis was then performed to classify the seventeen test bones into two categories: normal cases and outliers. This classification step depends on the measured parameters of the particular test bone. In case all parameters of a test bone were covered by the training population's parameter ranges, this bone is classified as normal bone, otherwise as outlier bone. Our experimental results showed that statistically there was no significant difference between the morphometric parameters extracted from the reconstructed surface models of the normal cases and those extracted from the reconstructed surface models of the outliers. Therefore, our statistical shape model based reconstruction technique can be used to reconstruct not only the surface model of a normal bone but also that of an outlier bone.

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Laser tissue soldering (LTS) is a promising technique for tissue fusion based on a heat-denaturation process of proteins. Thermal damage of the fused tissue during the laser procedure has always been an important and challenging problem. Particularly in LTS of arterial blood vessels strong heating of the endothelium should be avoided to minimize the risk of thrombosis. A precise knowledge of the temperature distribution within the vessel wall during laser irradiation is inevitable. The authors developed a finite element model (FEM) to simulate the temperature distribution within blood vessels during LTS. Temperature measurements were used to verify and calibrate the model. Different parameters such as laser power, solder absorption coefficient, thickness of the solder layer, cooling of the vessel and continuous vs. pulsed energy deposition were tested to elucidate their impact on the temperature distribution within the soldering joint in order to reduce the amount of further animal experiments. A pulsed irradiation with high laser power and high absorbing solder yields the best results.

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An algorithm for the real-time registration of a retinal video sequence captured with a scanning digital ophthalmoscope (SDO) to a retinal composite image is presented. This method is designed for a computer-assisted retinal laser photocoagulation system to compensate for retinal motion and hence enhance the accuracy, speed, and patient safety of retinal laser treatments. The procedure combines intensity and feature-based registration techniques. For the registration of an individual frame, the translational frame-to-frame motion between preceding and current frame is detected by normalized cross correlation. Next, vessel points on the current video frame are identified and an initial transformation estimate is constructed from the calculated translation vector and the quadratic registration matrix of the previous frame. The vessel points are then iteratively matched to the segmented vessel centerline of the composite image to refine the initial transformation and register the video frame to the composite image. Criteria for image quality and algorithm convergence are introduced, which assess the exclusion of single frames from the registration process and enable a loss of tracking signal if necessary. The algorithm was successfully applied to ten different video sequences recorded from patients. It revealed an average accuracy of 2.47 ± 2.0 pixels (∼23.2 ± 18.8 μm) for 2764 evaluated video frames and demonstrated that it meets the clinical requirements.

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Percutaneous needle intervention based on PET/CT images is effective, but exposes the patient to unnecessary radiation due to the increased number of CT scans required. Computer assisted intervention can reduce the number of scans, but requires handling, matching and visualization of two different datasets. While one dataset is used for target definition according to metabolism, the other is used for instrument guidance according to anatomical structures. No navigation systems capable of handling such data and performing PET/CT image-based procedures while following clinically approved protocols for oncologic percutaneous interventions are available. The need for such systems is emphasized in scenarios where the target can be located in different types of tissue such as bone and soft tissue. These two tissues require different clinical protocols for puncturing and may therefore give rise to different problems during the navigated intervention. Studies comparing the performance of navigated needle interventions targeting lesions located in these two types of tissue are not often found in the literature. Hence, this paper presents an optical navigation system for percutaneous needle interventions based on PET/CT images. The system provides viewers for guiding the physician to the target with real-time visualization of PET/CT datasets, and is able to handle targets located in both bone and soft tissue. The navigation system and the required clinical workflow were designed taking into consideration clinical protocols and requirements, and the system is thus operable by a single person, even during transition to the sterile phase. Both the system and the workflow were evaluated in an initial set of experiments simulating 41 lesions (23 located in bone tissue and 18 in soft tissue) in swine cadavers. We also measured and decomposed the overall system error into distinct error sources, which allowed for the identification of particularities involved in the process as well as highlighting the differences between bone and soft tissue punctures. An overall average error of 4.23 mm and 3.07 mm for bone and soft tissue punctures, respectively, demonstrated the feasibility of using this system for such interventions. The proposed system workflow was shown to be effective in separating the preparation from the sterile phase, as well as in keeping the system manageable by a single operator. Among the distinct sources of error, the user error based on the system accuracy (defined as the distance from the planned target to the actual needle tip) appeared to be the most significant. Bone punctures showed higher user error, whereas soft tissue punctures showed higher tissue deformation error.

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The excimer laser-assisted nonocclusive anastomosis (ELANA) technique has been developed as a clinical effective technique to perform intracranial high-flow bypass without temporary occlusion of cerebral vessels in otherwise untreatable or high-risk cerebrovascular diseases. We experimentally tested the application of a nonabsorbable cyanoacrylate-based sealant with the ELANA technique.

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The authors conducted an in vivo study to determine clinical cutoffs for a laser fluorescence (LF) device, an LF pen and a fluorescence camera (FC), as well as to evaluate the clinical performance of these methods and conventional methods in detecting occlusal caries in permanent teeth by using the histologic gold standard for total validation of the sample.

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A device based on infrared laser fluorescence (IRLF) has become available as an adjunct for the diagnosis of dental caries.

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OBJECTIVES: To assess influence of durotomy on spinal cord blood flow (SCBF) in chondrodystrophic dogs with thoracolumbar disk extrusion. STUDY DESIGN: Prospective cohort study. ANIMALS: Chondrodystrophic dogs with thoracolumbar disk extrusion (n = 11). METHODS: Diagnosis was based on neurologic signs, magnetic resonance imaging (MRI) findings, and surgical confirmation. Regional SCBF was measured 3 times intraoperatively by laser-Doppler flowmetry: (1) before surgical decompression; (2) immediately after decompression by hemilaminectomy-durotomy; and (3) after 15 minutes of lesion lavage. A standardized hemilaminectomy and durotomy performed by the same neurosurgeon, was used to minimize factors that could influence measurement readings. RESULTS: A significant increase in intraoperative SCBF was found immediately after spinal cord decompression and durotomy in dogs but SCBF returned to previous levels or lower after 15 minutes of lavage. Changes in SCBF were not associated with duration of clinical signs; neurologic status, degree of spinal cord compression, or signal intensity changes as assessed by MRI. CONCLUSION: Durotomy does not increase SCBF in dogs with disk extrusion associated spinal cord compression.

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Vitiligo is a relatively common acquired disorder, characterized by progressive loss of melanocytes from the epidermis and the epidermal appendages. The disease is associated with considerable morbidity because of a major impact on the quality of life. The treatment for vitiligo is generally unsatisfactory and challenging. There are a variety of therapeutic possibilities including topical corticosteroids, topical calcineurin inhibitors, as well as phototherapy with Psoralen plus UVA (PUVA), narrow-band UVB, and a 308-nm excimer laser and/or lamps. Furthermore, surgical methods encompass grafting and transplantation while depigmentation treatments and psychological support may also be considered. The objective is to assess the effect of the 380-nm excimer laser in the treatment of vitiligo based on the available studies and case series. We searched the relevant literature about vitiligo and excimer laser published between 1990 and 2012 using the MEDLINE database. We reviewed all relevant articles about 308-nm excimer laser and light sources assessing their efficacy in the management of vitiligo as well as their side effects. The value of combination treatment methods was also analyzed. The available studies provide strong evidence that the excimer laser represents the most effective approach to treat vitiligo compared to ordinary phototherapy. Excimer laser is relatively safe and effective for localized disease. UV-sensitive areas respond best as well as a short duration of the disease. More frequent treatments achieve better results. Compared to other treatment modalities, the excimer laser most likely constitutes the treatment of choice for localized vitiligo. Its efficacy can be further improved in combination with other therapies such as corticosteroids, pimecrolimus, or tacrolimus.

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This paper is meant to provide guidance to anyone wishing to write a neurological guideline for diagnosis or treatment, and is directed at the Scientist Panels and task forces of the European Federation of Neurological Societies (EFNS). It substitutes the previous guidance paper from 2004. It contains several new aspects: the guidance is now based on a change of the grading system for evidence and for the resulting recommendations, and has adopted The Grading of Recommendations, Assessment, Development and Evaluation system (GRADE). The process of grading the quality of evidence and strength of recommendations can now be improved and made more transparent. The task forces embarking on the development of a guideline must now make clearer and more transparent choices about outcomes considered most relevant when searching the literature and evaluating their findings. Thus, the outcomes chosen will be more critical, more patient-oriented and easier to translate into simple recommendations. This paper also provides updated practical recommendations for planning a guideline task force within the framework of the EFNS. Finally, this paper hopes to find the approval also by the relevant bodies of our future organization, the European Academy of Neurology.

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Traditionally, non-invasive monitoring of tidal volume in infants has been performed using impedance plethysmography analyzed using a one or two compartment model. We developed a new laser system for use in infants, which measures antero-posterior movement of the chest wall during quiet sleep. In 24 unsedated or sedated infants (11 healthy, 13 with respiratory disease), we examined whether the analysis of thoracoabdominal movement based on a three compartment model could more accurately estimate tidal volume in comparison to V(T) measured at the mouth. Using five laser signals, chest wall movements were measured at the right and left, upper and lower ribcage and the abdomen. Within the tidal volume range from 4.6 to 135.7 ml, a three compartment model showed good short term repeatability and the best agreement with tidal volume measured at mouth (r(2) = 0.86) compared to that of a single compartment model (r(2) = 0.62, P < 0.0001) and a two compartment model (r(2) = 0.82, P < 0.01), particularly in the presence of respiratory disease. Three compartment modeling of a 5 laser thoracoabdominal monitoring permits more accurate estimates of tidal volume in infants and potentially of regional differences of chest wall displacement in future studies.

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The new device DIAGNOdent pen based on red laser light induced fluorescence was introduced for the detection of approximal and occlusal caries. The aim of this study was to test its performance on occlusal surfaces. The new device comes with two different sapphire fibre tips: a cylindrical tip and a conical tip. The two new sapphire fibre tips were used and compared with the tip currently available with DIAGNOdent (DD). METHODS: The teeth were selected from a pool of extracted permanent human molars, which were stored frozen at -20 degrees C, until use. Prior to being measured the teeth were defrosted and cleaned. One hundred and nineteen teeth were selected and measured with the old tip and with the two new tips of the new device by two independent assessments. The teeth were histologically prepared and assessed for caries extension. RESULTS: Specificity values for D(1), D(2) and D(3) ranged between 0.69 and 0.89, sensitivity between 0.78 and 0.96. There were no statistically significant differences obtained between the two tips of the new and the one tip of the old device (p>0.05). Intra-examiner reliability with kappa values of >0.83 was high. CONCLUSIONS: In this study, the new laser fluorescence device performed on occlusal surfaces as well as the available device.

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Nitrous oxide fluxes were measured at the Lägeren CarboEurope IP flux site over the multi-species mixed forest dominated by European beech and Norway spruce. Measurements were carried out during a four-week period in October–November 2005 during leaf senescence. Fluxes were measured with a standard ultrasonic anemometer in combination with a quantum cascade laser absorption spectrometer that measured N2O, CO2, and H2O mixing ratios simultaneously at 5 Hz time resolution. To distinguish insignificant fluxes from significant ones it is proposed to use a new approach based on the significance of the correlation coefficient between vertical wind speed and mixing ratio fluctuations. This procedure eliminated roughly 56% of our half-hourly fluxes. Based on the remaining, quality checked N2O fluxes we quantified the mean efflux at 0.8±0.4 μmol m−2 h−1 (mean ± standard error). Most of the contribution to the N2O flux occurred during a 6.5-h period starting 4.5 h before each precipitation event. No relation with precipitation amount could be found. Visibility data representing fog density and duration at the site indicate that wetting of the canopy may have as strong an effect on N2O effluxes as does below-ground microbial activity. It is speculated that above-ground N2O production from the senescing leaves at high moisture (fog, drizzle, onset of precipitation event) may be responsible for part of the measured flux.