Comparison between laser scanning tomography and computerised image analysis of the optic disc

Aims - To study the interchangeability of the measurements of the optic disc topography obtained by one computerised image analyser and one confocal laser tomographic scanner. Methods - One eye of 28 patients with glaucoma or glaucoma suspects was studied. All cases had simultaneous stereoscopic disc photographs taken with the fundus camera Topcon TRC-SS and optic disc examination with the Heidelberg retina tomograph (HRT) during the same visit. The optic disc photographs were digitised and analysed with the Topcon ImageNet (TI) system. Three variables of the optic disc topography provided by the TI and the HRT were compared - cup volume (CV), rim area (RA), and cup area to disc area ratio (CA/DA). Results - The mean values of CV and RA provided by the TI (0.52 (SD 0.32) mm and 1.58 (0.39) mm , respectively) were greater (p <0.01) than the mean values of CV and RA determined by the HRT (0.32 (0.25) mm , and 1.33 (0.47) mm , respectively). The mean value of CA/DA provided by the TI (0.42 (0.14)) and the HRT (0.42 (0.18)) was similar (p = 0.93). Correlation coefficients between measurements obtained by the two methods ranged from 0.53 to 0.73. Conclusion - There was a significant discrepancy in the measurements of rim area and cup volume of the optic disc obtained by a computerised image analyser and a laser scanning tomograph.

HD Laser scanning for the evaluation of salt decay laboratory simulations of building limestone

Laboratory salt decay simulations are a well established method to assess the relative durability of stone. There is still, however, very much scope to implement improved monitoring techniques to investigate the changes experienced by the materials during these experiments. Non-destructive techniques have acquired over recent decades a preferential status for monitoring change samples during salt decay tests, as they allow cumulative tests on each sample. The development of HD laser scanning permits detailed mapping of surface changes and, therefore, constitutes an effective technique to monitor non-destructively surface changes in tested samples as an alternative to other monitoring techniques such as traditional weight loss strategies that do not permit any degree of spatial differentiation that can be related, for example, to underlying stone properties.

Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.

Red blood cell (RBC) parameters such as morphology, volume, refractive index, and hemoglobin content are of great importance for diagnostic purposes. Existing approaches require complicated calibration procedures and robust cell perturbation. As a result, reference values for normal RBC differ depending on the method used. We present a way for measuring parameters of intact individual RBCs by using digital holographic microscopy (DHM), a new interferometric and label-free technique with nanometric axial sensitivity. The results are compared with values achieved by conventional techniques for RBC of the same donor and previously published figures. A DHM equipped with a laser diode (lambda = 663 nm) was used to record holograms in an off-axis geometry. Measurements of both RBC refractive indices and volumes were achieved via monitoring the quantitative phase map of RBC by means of a sequential perfusion of two isotonic solutions with different refractive indices obtained by the use of Nycodenz (decoupling procedure). Volume of RBCs labeled by membrane dye Dil was analyzed by confocal microscopy. The mean cell volume (MCV), red blood cell distribution width (RDW), and mean cell hemoglobin concentration (MCHC) were also measured with an impedance volume analyzer. DHM yielded RBC refractive index n = 1.418 +/- 0.012, volume 83 +/- 14 fl, MCH = 29.9 pg, and MCHC 362 +/- 40 g/l. Erythrocyte MCV, MCH, and MCHC achieved by an impedance volume analyzer were 82 fl, 28.6 pg, and 349 g/l, respectively. Confocal microscopy yielded 91 +/- 17 fl for RBC volume. In conclusion, DHM in combination with a decoupling procedure allows measuring noninvasively volume, refractive index, and hemoglobin content of single-living RBCs with a high accuracy.

Bone growth around silicon nitride implants - An evaluation by scanning electron microscopy

Silicon nitride has demonstrated to be a potential candidate for clinical applications because it is a non-cytotoxic material and has satisfactory fracture toughness, high wear resistance and low friction coefficient. In this paper, samples of silicon nitride, which were kept into rabbits` tibias for 8 weeks, and the adjacentbone tissue were analysed by scanning electron microscopy in order to verify the bone growth around the implants and the interaction between the implant and the bone. Bone growth occurred mainly in the cortical areas, although it has been observed that the newly bone tends to grow toward the marrow cavity. Differences were observed between the implants installed into distal and proximal regions. In the first region, where the distance between the implant and the cortical bone is greater than in the proximal region, the osteoconduction process was evidenced by the presence of a bridge bone formation toward the implant surface. The results showed that silicon nitride can be used as biomaterial since the newly bone grew around the implants. (c) 2007 Elsevier Inc. All rights reserved.

Study of the biodegradation of a polymer derived from castor oil by scanning electron microscopy, thermogravimetry and infrared spectroscopy

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Scanning electron microscopy evaluation of the effect of etching agents on human enamel surface

Acid etching promotes microporosities on enamel surface, which provide a better bonding surface to adhesive materials. The purpose of this study was to comparatively analyze the microstructure of enamel surface after etching with 37% phosphoric acid or with two self-etching primers, Non-rinse conditioner (NRC) and Clearfil SE Bond (CSEB) using scanning electron microscopy. Thirty sound premolars were divided into 3 groups with ten teeth each: Group 1: the buccal surface was etched with 37% phosphoric acid for 15 seconds; Group 2: the buccal surface was etched with NRC for 20 seconds; Group 3: the buccal surface was etched with CSEB for 20 seconds. Teeth from Group 1 were rinsed with water; teeth from all groups were air-dried for 15 seconds. After that, all specimens were processed for scanning electron microscopy and analyzed in a Jeol 6100 SEM. The results showed deeper etching when the enamel surface was etched with 37% phosphoric acid, followed by NRC and CSEB. It is concluded that 37% phosphoric acid is still the best agent for a most effective enamel etching.

Evaluation of early changes induced by diuron in the rat urinary bladder using different processing methods for scanning electron microscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of the bleached human enamel by scanning electron microscopy

Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning Electron Microscopy (SEM). Materials and Methods: Twenty intact human third molars extracted for orthodontic reasons were randomly divided into four groups (n=5) treated as follows: G1- storage in artificial saliva (control group); G2- four 30-minute applications of 35% carbamide peroxide (total exposure: 2h); G3- four 2-hour exposures to 35% carbamide peroxide (total exposure: 8h); G4- two applications of 35% hydrogen peroxide, which was light-activated with halogen lamp at 700mW/cm2 during 7min and remained in contact with enamel for 20min (total exposure: 40min). All bleaching treatments adopted in this study followed the application protocols advised by manufacturers. Evaluation of groups submitted to 35% carbamide peroxide was carried out after two time intervals (30 minutes and 2 hours per session), following the extreme situations recommended by the manufacturer. Specimens were prepared for SEM analysis performing gold sputter coating under vacuum and were examined using 15kV at 500x and 2000x magnification. Results: Morphological alterations on the enamel surface were similarly detected after bleaching with either 35% carbamide peroxide or 35% hydrogen peroxide. Surface porosities were characteristic of an erosive process that took place on human enamel. Depression areas, including the formation of craters, and exposure of enamel rods could also be detected. Conclusion: Bleaching effects on enamel morphology were randomly distributed throughout enamel surface and various degrees of enamel damage could be noticed. Clinical significance: In-office bleaching materials may adversely affect enamel morphology and therefore should be used with caution.

Avaliação dos fatores de virulência, atividade antimicrobiana e viabilidade celular de bactéria cariogênica na presença do Terpinen-4-ol: estudo in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos fatores de virulência, atividade antimicrobiana e viabilidade celular de bactéria cariogênica na presença do Terpinen-4-ol: estudo in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Probing the Enamel Topography After Acid Erosion by Scanning Electrochemical Microscopy

In this work, we present an investigation on the thickness of the eroded enamel layer in tooth samples after exposure to citric and hydrochloric acid by using Scanning Electrochemical Microscopy (SECM). Approaching curves with typical negative feedback behavior were obtained in enamel samples for evaluation of topographic changes. In a control experiment, SECM images showed no significant difference in the current monitored during the scan, implying that enamel demineralization did not occur in mineral water medium. Topographic SECM images obtained after contact with citric and hydrochloric acid for different periods of time showed a significant increase in the current relative to a previously protected surface, indicating the structural loss of enamel. The thickness of the enamel layer eroded after contact with hydrochloric acid was significantly higher when compared to the one obtained with citric acid. Hence, our results showed that the enamel acid erosion is a relatively fast process, which is strongly dependent on parameters such as pH, time, acid strength and acid concentration.

Connecting accuracy of interchanged prosthetic abutments to different dental implants using scanning electron microscopy

The present study evaluated the interchangeability of prosthetic components for external hexagon implants by measuring the precision of the implant/abutment (I/A) interface with scanning electron microscopy. Ten implants for each of three brands (SIN, Conexão, Neodent) were tested with their respective abutments (milled CoCr collar rotational and non-rotational) and another of an alternative manufacturer (Microplant) in randomly arranged I/A combinations. The degree of interchangeability between the various brands of components was defined using the original abutment interface gap with its respective implant as the benchmark dimension. Accordingly, when the result for a given component placed on an implant was equal to or smaller then that gap measured when the original component of the same brand as the implant was positioned, interchangeability was considered valid. Data were compared with the Kruskal-Wallis test at 5% significance level. Some degree of misfit was observed in all specimens. Generally, the non-rotational component was more accurate than its rotational counterpart. The latter samples ranged from 0.6-16.9 µm, with a 4.6 µm median; and the former from 0.3-12.9 µm, with a 3.4 µm median. Specimens with the abutment and fixture from Conexão had larger microgap than the original set for SIN and Neodent (p<0.05). Even though the latter systems had similar results with their respective components, their interchanged abutments did not reproduce the original accuracy. The results suggest that the alternative brand abutment would have compatibility with all systems while the other brands were not completely interchangeable.

Nanoscopic studies of conjugated polymer blends by (electric) scanning probe microscopy

Conjugated polymers and conjugated polymer blends have attracted great interest due to their potential applications in biosensors and organic electronics. The sub-100 nm morphology of these materials is known to heavily influence their electromechanical properties and the performance of devices they are part of. Electromechanical properties include charge injection, transport, recombination, and trapping, the phase behavior and the mechanical robustness of polymers and blends. Electrical scanning probe microscopy techniques are ideal tools to measure simultaneously electric (conductivity and surface potential) and dielectric (dielectric constant) properties, surface morphology, and mechanical properties of thin films of conjugated polymers and their blends.rnIn this thesis, I first present a combined topography, Kelvin probe force microscopy (KPFM), and scanning conductive torsion mode microscopy (SCTMM) study on a gold/polystyrene model system. This system is a mimic for conjugated polymer blends where conductive domains (gold nanoparticles) are embedded in a non-conductive matrix (polystyrene film), like for polypyrrole:polystyrene sulfonate (PPy:PSS), and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS). I controlled the nanoscale morphology of the model by varying the distribution of gold nanoparticles in the polystyrene films. I studied the influence of different morphologies on the surface potential measured by KPFM and on the conductivity measured by SCTMM. By the knowledge I gained from analyzing the data of the model system I was able to predict the nanostructure of a homemade PPy:PSS blend.rnThe morphologic, electric, and dielectric properties of water based conjugated polymer blends, e.g. PPy:PSS or PEDOT:PSS, are known to be influenced by their water content. These properties also influence the macroscopic performance when the polymer blends are employed in a device. In the second part I therefore present an in situ humidity-dependence study on PPy:PSS films spin-coated and drop-coated on hydrophobic highly ordered pyrolytic graphite substrates by KPFM. I additionally used a particular KPFM mode that detects the second harmonic electrostatic force. With this, I obtained images of dielectric constants of samples. Upon increasing relative humidity, the surface morphology and composition of the films changed. I also observed that relative humidity affected thermally unannealed and annealed PPy:PSS films differently. rnThe conductivity of a conjugated polymer may change once it is embedded in a non-conductive matrix, like for PPy embedded in PSS. To measure the conductivity of single conjugated polymer particles, in the third part, I present a direct method based on microscopic four-point probes. I started with metal core-shell and metal bulk particles as models, and measured their conductivities. The study could be extended to measure conductivity of single PPy particles (core-shell and bulk) with a diameter of a few micrometers.

Studies of degradation of organic solar cell materials using electrical scanning probe microscopy

Intense research is being done in the field of organic photovoltaics in order to synthesize low band-gap organic molecules. These molecules are electron donors which feature in combination with acceptor molecules, typically fullerene derivarntives, forming an active blend. This active blend has phase separated bicontinuous morphology on a nanometer scale. The highest recorded power conversionrnefficiencies for such cells have been 10.6%. Organic semiconductors differ from inorganic ones due to the presence of tightly bonded excitons (electron-hole pairs)resulting from their low dielectric constant (εr ≈2-4). An additional driving force is required to separate such Frenkel excitons since their binding energy (0.3-1 eV) is too large to be dissociated by an electric field alone. This additional driving force arises from the energy difference between the lowest unoccupied molecular orbital (LUMO) of the donor and the acceptor materials. Moreover, the efficiency of the cells also depends on the difference between the highest occupied molecular orbital (HOMO) of the donor and LUMO of the acceptor. Therefore, a precise control and estimation of these energy levels are required. Furthermore any external influences that change the energy levels will cause a degradation of the power conversion efficiency of organic solar cell materials. In particular, the role of photo-induced degradation on the morphology and electrical performance is a major contribution to degradation and needs to be understood on a nanometer scale. Scanning Probe Microscopy (SPM) offers the resolution to image the nanometer scale bicontinuous morphology. In addition SPM can be operated to measure the local contact potential difference (CPD) of materials from which energy levels in the materials can be derived. Thus SPM is an unique method for the characterization of surface morphology, potential changes and conductivity changes under operating conditions. In the present work, I describe investigations of organic photovoltaic materials upon photo-oxidation which is one of the major causes of degradation of these solar cell materials. SPM, Nuclear Magnetic Resonance (NMR) and UV-Vis spectroscopy studies allowed me to identify the chemical reactions occurring inside the active layer upon photo-oxidation. From the measured data, it was possible to deduce the energy levels and explain the various shifts which gave a better understanding of the physics of the device. In addition, I was able to quantify the degradation by correlating the local changes in the CPD and conductivity to the device characteristics, i.e., open circuit voltage and short circuit current. Furthermore, time-resolved electrostatic force microscopy (tr-EFM) allowed us to probe dynamic processes like the charging rate of the individual donor and acceptor domains within the active blend. Upon photo-oxidation, it was observed, that the acceptor molecules got oxidized first preventing the donor polymer from degrading. Work functions of electrodes can be tailored by modifying the interface with monomolecular thin layers of molecules which are made by a chemical reaction in liquids. These modifications in the work function are particularly attractive for opto-electronic devices whose performance depends on the band alignment between the electrodes and the active material. In order to measure the shift in work function on a nanometer scale, I used KPFM in situ, which means in liquids, to follow changes in the work function of Au upon hexadecanethiol adsorption from decane. All the above investigations give us a better understanding of the photo-degradation processes of the active material at the nanoscale. Also, a method to compare various new materials used for organic solar cells for stability is proposed which eliminates the requirement to make fully functional devices saving time and additional engineering efforts.

Wet and dry model granulates under mechanical load : a confocal microscopy study

Granular matter, also known as bulk solids, consists of discrete particles with sizes between micrometers and meters. They are present in many industrial applications as well as daily life, like in food processing, pharmaceutics or in the oil and mining industry. When handling granular matter the bulk solids are stored, mixed, conveyed or filtered. These techniques are based on observations in macroscopic experiments, i.e. rheological examinations of the bulk properties. Despite the amply investigations of bulk mechanics, the relation between single particle motion and macroscopic behavior is still not well understood. For exploring the microscopic properties on a single particle level, 3D imaging techniques are required.rnThe objective of this work was the investigation of single particle motions in a bulk system in 3D under an external mechanical load, i.e. compression and shear. During the mechanical load the structural and dynamical properties of these systems were examined with confocal microscopy. Therefor new granular model systems in the wet and dry state were designed and prepared. As the particles are solid bodies, their motion is described by six degrees of freedom. To explore their entire motion with all degrees of freedom, a technique to visualize the rotation of spherical micrometer sized particles in 3D was developed. rnOne of the foci during this dissertation was a model system for dry cohesive granular matter. In such systems the particle motion during a compression of the granular matter was investigated. In general the rotation of single particles was the more sensitive parameter compared to the translation. In regions with large structural changes the rotation had an earlier onset than the translation. In granular systems under shear, shear dilatation and shear zone formation were observed. Globally the granular sediments showed a shear behavior, which was known already from classical shear experiments, for example with Jenike cells. Locally the shear zone formation was enhanced, when near the applied load a pre-diluted region existed. In regions with constant volume fraction a mixing between the different particle layers occurred. In particular an exchange of particles between the current flowing region and the non-flowing region was observed. rnThe second focus was on model systems for wet granular matter, where an additional binding liquid is added to the particle suspension. To examine the 3D structure of the binding liquid on the micrometer scale independently from the particles, a second illumination and detection beam path was implemented. In shear and compression experiments of wet clusters and bulk systems completely different dynamics compared to dry cohesive models systems occured. In a Pickering emulsion-like system large structural changes predominantly occurred in the local environment of binding liquid droplets. These large local structural changes were due to an energy interplay between the energy stored in the binding droplet during its deformation and the binding energy of particles at the droplet interface. rnConfocal microscopy in combination with nanoindentation gave new insights into the single particle motions and dynamics of granular systems under a mechanical load. These novel experimental results can help to improve the understanding of the relationship between bulk properties of granular matter, such as volume fraction or yield stress and the dynamics on a single particle level.rnrn