Fluorescence lifetime imaging microscopy (FLIM) to demonstrate the nuclear binding of flavanols and (--epigallocatechin gallate

The use of light microscopy and DMACA staining strongly suggested that plant and animal cell nuclei act as sinks for flavanols [1, 2]. Detailed uv-vis spectroscopic titration experiments indicated that histone proteins are the likely binding sites in the nucleus [2]. Here we report the development of a multi-photon excitation microscopy technique combined with fluorescent lifetime measurements of flavanols. Using this technique, (+) catechin, (-) epicatechin and (-) epigallocatechin gallate (EGCG) showed strikingly different excited state lifetimes in solution. Interaction of histone proteins with flavanols was indicated by the appearance of a significant τ2-component of 1.7 to 4.0ns. Tryptophan interference could be circumvented in the in vivo fluorescence lifetime imaging microscopy (FLIM) experiments with 2-photon excitation at 630nm. This enabled visualisation and semi-quantitative measurements that demonstrated unequivocally the absorption of (+)catechin, (-)epicatechin and EGCG by nuclei of onion cells. 3D FLIM revealed for the first time that externally added EGCG penetrated the whole nucleus in onion cells. The relative proportions of EGCG in cytoplasm: nucleus: nucleoli were ca. 1:10:100. FLIM experiments may therefore facilitate probing the health effects of EGCG, which is the major constituent of green tea.

A 3D cloud-construction algorithm for the EarthCARE satellite mission

This article presents and assesses an algorithm that constructs 3D distributions of cloud from passive satellite imagery and collocated 2D nadir profiles of cloud properties inferred synergistically from lidar, cloud radar and imager data. It effectively widens the active–passive retrieved cross-section (RXS) of cloud properties, thereby enabling computation of radiative fluxes and radiances that can be compared with measured values in an attempt to perform radiative closure experiments that aim to assess the RXS. For this introductory study, A-train data were used to verify the scene-construction algorithm and only 1D radiative transfer calculations were performed. The construction algorithm fills off-RXS recipient pixels by computing sums of squared differences (a cost function F) between their spectral radiances and those of potential donor pixels/columns on the RXS. Of the RXS pixels with F lower than a certain value, the one with the smallest Euclidean distance to the recipient pixel is designated as the donor, and its retrieved cloud properties and other attributes such as 1D radiative heating rates are consigned to the recipient. It is shown that both the RXS itself and Moderate Resolution Imaging Spectroradiometer (MODIS) imagery can be reconstructed extremely well using just visible and thermal infrared channels. Suitable donors usually lie within 10 km of the recipient. RXSs and their associated radiative heating profiles are reconstructed best for extensive planar clouds and less reliably for broken convective clouds. Domain-average 1D broadband radiative fluxes at the top of theatmosphere(TOA)for (21 km)2 domains constructed from MODIS, CloudSat andCloud–Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) data agree well with coincidental values derived from Clouds and the Earth’s Radiant Energy System (CERES) radiances: differences betweenmodelled and measured reflected shortwave fluxes are within±10Wm−2 for∼35% of the several hundred domains constructed for eight orbits. Correspondingly, for outgoing longwave radiation∼65% are within ±10Wm−2.

3D digital elevation model generation

This chapter presents techniques used for the generation of 3D digital elevation models (DEMs) from remotely sensed data. Three methods are explored and discussed—optical stereoscopic imagery, Interferometric Synthetic Aperture Radar (InSAR), and LIght Detection and Ranging (LIDAR). For each approach, the state-of-the-art presented in the literature is reviewed. Techniques involved in DEM generation are presented with accuracy evaluation. Results of DEMs reconstructed from remotely sensed data are illustrated. While the processes of DEM generation from satellite stereoscopic imagery represents a good example of passive, multi-view imaging technology, discussed in Chap. 2 of this book, InSAR and LIDAR use different principles to acquire 3D information. With regard to InSAR and LIDAR, detailed discussions are conducted in order to convey the fundamentals of both technologies.

Subdiffraction fluorescence imaging of biomolecular structure and distributions with quantum dots

We introduce semiconductor quantum dot-based fluorescence imaging with approximately 2-fold increased optical resolution in three dimensions as a method that allows both studying cellular structures and spatial organization of biomolecules in membranes and subcellular organelles. Target biomolecules are labelled with quantum dots via immunocytochemistry. The resolution enhancement is achieved by three-photon absorption of quantum dots and subsequent fluorescence emission from a higher-order excitonic state. Different from conventional multiphoton microscopy, this approach can be realized on any confocal microscope without the need for pulsed excitation light. We demonstrate quantum dot triexciton imaging (QDTI) of the microtubule network of U373 cells, 3D imaging of TNF receptor 2 on the plasma membrane of HeLa cells, and multicolor 3D imaging of mitochondrial cytochrome c oxidase and actin in COS-7 cells.

Simple method for sub-diffraction resolution imaging of cellular structures on standard confocal microscopes by three-photon absorption of quantum dots

This study describes a simple technique that improves a recently developed 3D sub-diffraction imaging method based on three-photon absorption of commercially available quantum dots. The method combines imaging of biological samples via tri-exciton generation in quantum dots with deconvolution and spectral multiplexing, resulting in a novel approach for multi-color imaging of even thick biological samples at a 1.4 to 1.9-fold better spatial resolution. This approach is realized on a conventional confocal microscope equipped with standard continuous-wave lasers. We demonstrate the potential of multi-color tri-exciton imaging of quantum dots combined with deconvolution on viral vesicles in lentivirally transduced cells as well as intermediate filaments in three-dimensional clusters of mouse-derived neural stem cells (neurospheres) and dense microtubuli arrays in myotubes formed by stacks of differentiated C2C12 myoblasts.

3D Cell-Migration Studies using Two-Photon Engineered Polymer Scaffolds

We use two-photon polymerization to fabricate 3D scaffolds with precise control over pore size and shape for studying cell migration in 3D. These scaffolds allow movement of cells in all directions. The fabrication, imaging, and quantitative analysis method developed here can be used to do systematic cell studies in 3D.

Refraction corrected transmission ultrasound computed tomography for application in breast imaging

Purpose: We present an iterative framework for CT reconstruction from transmission ultrasound data which accurately and efficiently models the strong refraction effects that occur in our target application: Imaging the female breast. Methods: Our refractive ray tracing framework has its foundation in the fast marching method (FNMM) and it allows an accurate as well as efficient modeling of curved rays. We also describe a novel regularization scheme that yields further significant reconstruction quality improvements. A final contribution is the development of a realistic anthropomorphic digital breast phantom based on the NIH Visible Female data set. Results: Our system is able to resolve very fine details even in the presence of significant noise, and it reconstructs both sound speed and attenuation data. Excellent correspondence with a traditional, but significantly more computationally expensive wave equation solver is achieved. Conclusions: Apart from the accurate modeling of curved rays, decisive factors have also been our regularization scheme and the high-quality interpolation filter we have used. An added benefit of our framework is that it accelerates well on GPUs where we have shown that clinical 3D reconstruction speeds on the order of minutes are possible.

Survey radiography and computerized tomography imaging of the thorax in female dogs with mammary tumors

Background: Accurate early diagnosis of lung metastases is important for establishing therapeutic measures. Therefore, the present study aimed to compare survey thoracic radiographs and computerized tomography (CT) scans to specifically identify lung metastases in female dogs with mammary tumors.Methods: Twenty-one female dogs, weighing 3 to 34 kg and aged from 5 years to 14 years and 10 months, with mammary tumors were studied. In all dogs before the imaging examinations, fine-needle aspiration cytology of the mammary tumors was performed to confirm the diagnosis. Three-view thoracic radiographs were accomplished: right lateral, left lateral and ventrodorsal views. Sequential transverse images of the thorax were acquired on a spiral Scanner, before and after intravenous bolus injection of nonionic iodine contrast. Soft-tissue and lung windows were applied. All the mammary tumors were surgically removed and examined histologically.Results: The correlation between the cytological and histological results regarding presence of malignancy was observed in only 17 cases. In radiographic examinations, no dog displayed signs of lung metastases or thorax chest lesions. CT detected lung metastasis in two cases, while small areas of lung atelectasis located peripherally were found in 28.57% of the dogs.Conclusion: In this study population, spiral CT showed higher sensitivity than chest radiographies to detect lung metastasis; this indicates that CT should be performed on all female dogs with malignant mammary tumors.

Association of 3D Reconstruction and Conventional Radiography for the Description of the Appendicular Skeleton of Chelonoidis carbonaria (Spix, 1824)

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

Evaluation of Different Methods of Optical Impression Making on the Marginal Gap of Onlays Created with CEREC 3D

Objectives: This study evaluated the marginal gaps on several surfaces of onlays created with the Cerec 3D system using one intraoral and two extraoral optical impression methods. Methods: A human molar (#19) was mounted with its adjacent teeth on a typodont (Frasaco) and prepared for a MODL onlay. The typodont was assembled in the mannequin head in order to simulate clinical conditions. The same operator took 36 individual optical impressions using a CEREC 3D camera. For group 1 (IP), a thin layer of titanium dioxide powder (CEREC powder-VITA) was applied directly onto the surface of the preparation for imaging (n=12). For group 2 (EP), a sectional impression was taken with hydrocolloid Identic Syringable (Dux Dental), a die made with polyvinylsiloxane KwikkModel Scan (R-dental Dentalerzeugnisse GmbH) and powdered with titanium dioxide for imaging (n=12). For group 3 (ES), a sectional impression was taken with PVS and a sectional stock tray, a die fabricated in stone (Diamond die- HI-TEC Dental Products) and the die being imaged without powdering (n=12). One operator designed and machined the onlays in Vita Blocks Mark II for Cerec (VITA) using a CEREC 3D. The marginal gaps (pm) were measured with an optical microscope (50x) at 12 points, three on each surface of the MODL. The results were analyzed by two-way ANOVA/Tukey's (p=0.05). Results: The overall mean marginal gaps (mu m) for the three methods were: IP=111.6 (+/- 34.0); EP=161.4 (+/- 37.6) and ES=116.8 (+/- 42.3). IP and ES were equal, but both were significantly less than EP. The pooled mean marginal gaps (mu m) for the occlusal = 110.5 (+/- 39) and lingual = 111.5 (+/- 30.5) surfaces were equivalent and significantly less than the distal = 136.5 (+/- 42.5) and mesial = 161.1 (+/- 43.3). Conclusion: The marginal gap of CEREC 3D onlay restorations was not different when the optical impression was taken intraorally vs extraorally using a stone cast that does not require powdering. The lingual and occlusal surfaces showed the lowest gaps.

Using GPUs for beamforming acceleration on SAFT imaging

SAFT techniques are based on the sequential activation, in emission and reception, of the array elements and the post-processing of all the received signals to compose the image. Thus, the image generation can be divided into two stages: (1) the excitation and acquisition stage, where the signals received by each element or group of elements are stored; and (2) the beamforming stage, where the signals are combined together to obtain the image pixels. The use of Graphics Processing Units (GPUs), which are programmable devices with a high level of parallelism, can accelerate the computations of the beamforming process, that usually includes different functions such as dynamic focusing, band-pass filtering, spatial filtering or envelope detection. This work shows that using GPU technology can accelerate, in more than one order of magnitude with respect to CPU implementations, the beamforming and post-processing algorithms in SAFT imaging. ©2009 IEEE.

Bone apposition to laminin-1 coated implants: Histologic and 3D evaluation

Laminin-1 has been reported as one of the factors responsible for the nucleation of calcium phosphates and, in vitro, has been reported to selectively recruit osteoprogenitors. This article focused on its in vivo effects, and evaluated the effect of laminin-1 local application on osseointegration. Polished cylindrical hydroxyapatite implants were coated with laminin-1 (test) and the bone responses in the rabbit tibiae after 2 and 4 weeks were evaluated and compared to the non-coated implants (control). Before the samples were processed for histological sectioning, they were three-dimensionally analysed with micro computed tomography (μCT). Both evaluation methods were analysed with regards to bone area around the implant and bone to implant contact. From the histologic observation, new bone formation around the laminin-1 coated implant at 2 weeks seemed to have increased the amount of supporting bone around the implant, however, at 4 weeks, the two groups presented no notable differences. The two-dimensional and three-dimensional morphometric evaluation revealed that both histologic and three-dimensional analysis showed some tendency in favour of the test group implants, however there was no statistical significance between the test and control group results. © 2012 International Association of Oral and Maxillofacial Surgeons.

Free tools and strategies for the generation of 3D finite element meshes: Modeling of the cardiac structures

The Finite Element Method is a well-known technique, being extensively applied in different areas. Studies using the Finite Element Method (FEM) are targeted to improve cardiac ablation procedures. For such simulations, the finite element meshes should consider the size and histological features of the target structures. However, it is possible to verify that some methods or tools used to generate meshes of human body structures are still limited, due to nondetailed models, nontrivial preprocessing, or mainly limitation in the use condition. In this paper, alternatives are demonstrated to solid modeling and automatic generation of highly refined tetrahedral meshes, with quality compatible with other studies focused on mesh generation. The innovations presented here are strategies to integrate Open Source Software (OSS). The chosen techniques and strategies are presented and discussed, considering cardiac structures as a first application context. © 2013 E. Pavarino et al.

Tilted and Short Implants Supporting Fixed Prosthesis in an Atrophic Maxilla: A 3D-FEA Biomechanical Evaluation

Purpose: This study compared the biomechanical behavior of tilted long implant and vertical short implants to support fixed prosthesis in an atrophic maxilla. Materials and Methods: The maxilla model was built based on a tomographic image of the patient. Implant models were based on micro-computer tomography imaging of implants. The different configurations considered were M4S, four vertical anterior implants; M4T, two mesial vertical implants and two distal tilted (45°) implants in the anterior region of the maxilla; and M6S, four vertical anterior implants and two vertical posterior implants. Numerical simulation was carried out under bilateral 150N loads applied in the cantilever region in axial (L1) and oblique (45°) (L2) direction. Bone was analyzed using the maximum and minimum principal stress (σmax and σmin), and von Mises stress (σvM) assessments. Implants were analyzed using the σvM. Results: The higher σmax was observed at: M4T, followed by M6S/L1, M6S/L2, M4S/L2, and M4S/L1 and the higher σvM: M4T/L1, M4T/L2 and M4S/L2, M6S/L2, M4S/L1, and M6S/L1. Conclusions: The presence of distal tilted (all-on-four) and distal short implants (all-on-six) resulted in higher stresses in both situations in the maxillary bone in comparison to the presence of vertical implants (all-on-four). © 2013 Wiley Periodicals, Inc.

Aspectos históricos e implicações da utilização do efeito 3D no cinema: o caso de A invenção de Hugo Cabret

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)