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.

An Alternative Section Method for Casting and Posterior Laser Welding of Metallic Frameworks for an Implant-Supported Prosthesis

Purpose: The aim of this study was to compare the accuracy of fit of three types of implant-supported frameworks cast in Ni-Cr alloy: specifically, a framework cast as one piece compared to frameworks cast separately in sections to the transverse or the diagonal axis, and later laser welded. Materials and Methods: Three sets of similar implant-supported frameworks were constructed. The first group of six 3-unit implant-supported frameworks were cast as one piece, the second group of six were sectioned in the transverse axis of the pontic region prior to casting, and the last group of six were sectioned in the diagonal axis of the pontic region prior to casting. The sectioned frameworks were positioned in the matrix (10 N(.)cm torque) and laser welded. To evaluate passive fit, readings were made with an optical microscope with both screws tightened and with only one-screw tightened. Data were submitted to ANOVA and Tukey-Kramer`s test (p < 0.05). Results: When both screws were tightened, no differences were found between the three groups (p > 0.05). In the single-screw-tightened test, with readings made opposite to the tightened side, the group cast as one piece (57.02 +/- 33.48 mu m) was significantly different (p < 0.05) from the group sectioned diagonally (18.92 +/- 4.75 mu m) but no different (p > 0.05) from the group transversally sectioned (31.42 +/- 20.68 mu m). On the tightened side, no significant differences were found between the groups (p > 0.05). Conclusions: Results of this study showed that casting diagonally sectioned frameworks lowers misfit levels of prosthetic implant-supported frameworks and also improves the levels of passivity to the same frameworks when compared to structures cast as one piece.

A scanning electron microscopy study of diseased root surfaces conditioned with EDTA gel plus Cetavlon after scaling and root planing

In the present investigation, a scanning electron microscopy analysis was performed to evaluate the effects of the topical application of ethylenediaminetetraacetic acid (EDTA) gel associated with Cetavlon (EDTAC) in removing the smear layer and exposing collagen fibers following root surface instrumentation. Twenty-eight teeth from adult humans, single rooted and scheduled for extraction due to periodontal reasons, were selected. Each tooth was submitted to manual (scaling and root planing) instrumentation alone or combined with ultrasonic instruments, with or without etching using a 24% EDTAC gel. Following extraction, specimens were processed and examined under a scanning electron microscope. A comparative morphological semi-quantitative analysis was performed; the intensity of the smear layer and the decalcification of cementum and dentinal surfaces were graded in 12 sets using an arbitrary scale ranging from 1 (area covered by a smear layer) to 4 (no smear layer). Root debridement with hand instruments alone or combined with ultrasonic instruments resulted in a similar smear layer covering the root surfaces. The smear layer was successfully removed from the surfaces treated with EDTAC, which exhibited numerous exposed dentinal tubules and collagen fibers. This study supports the hypothesis that manual instrumentation alone or instrumentation combined with ultrasonic instrumentation is unable to remove the smear layer, whereas the subsequent topical application of EDTAC gel effectively removes the smear layer, uncovers dentinal openings and exposes collagen fibers.

Modified section method for laser-welding of ill-fitting cp Ti and Ni-Cr alloy one-piece cast implant-supported frameworks

P>This study aimed to verify the effect of modified section method and laser-welding on the accuracy of fit of ill-fitting commercially pure titanium (cp Ti) and Ni-Cr alloy one-piece cast frameworks. Two sets of similar implant-supported frameworks were constructed. Both groups of six 3-unit implant-supported fixed partial dentures were cast as one-piece [I: Ni-Cr (control) and II: cp Ti] and evaluated for passive fitting in an optical microscope with both screws tightened and with only one screw tightened. All frameworks were then sectioned in the diagonal axis at the pontic region (III: Ni-Cr and IV: cp Ti). Sectioned frameworks were positioned in the matrix (10-Ncm torque) and laser-welded. Passive fitting was evaluated for the second time. Data were submitted to anova and Tukey-Kramer honestly significant difference tests (P < 0 center dot 05). With both screws tightened, one-piece cp Ti group II showed significantly higher misfit values (27 center dot 57 +/- 5 center dot 06 mu m) than other groups (I: 11 center dot 19 +/- 2 center dot 54 mu m, III: 12 center dot 88 +/- 2 center dot 93 mu m, IV: 13 center dot 77 +/- 1 center dot 51 mu m) (P < 0 center dot 05). In the single-screw-tightened test, with readings on the opposite side to the tightened side, Ni-Cr cast as one-piece (I: 58 center dot 66 +/- 14 center dot 30 mu m) was significantly different from cp Ti group after diagonal section (IV: 27 center dot 51 +/- 8 center dot 28 mu m) (P < 0 center dot 05). On the tightened side, no significant differences were found between groups (P > 0 center dot 05). Results showed that diagonally sectioning ill-fitting cp Ti frameworks lowers misfit levels of prosthetic implant-supported frameworks and also improves passivity levels of the same frameworks when compared to one-piece cast structures.

A light and scanning electron microscopy study of bone healing following inferior alveolar nerve lateralization: An experimental study in rabbits

Purpose: The purpose of this study was to evaluate the bone healing kinetics around commercially pure titanium implants following inferior alveolar nerve (IAN) lateralization in a rabbit model. Materials and Methods: Inferior alveolar nerve lateralization was performed in 16 adult female rabbits (Oryctolagus cuniculus). During the nerve lateralization procedure, 1 implant was placed through the mandibular canal, and the IAN was replaced in direct contact with the implant. During the 8-week healing period, various bone labels were administered for fluorescent microscopy analysis. The animals were euthanized by anesthesia overdose, and the mandibular blocks were exposed by sharp dissection. Nondecalcified samples were prepared for optical light and scanning electron microscopy (SEM) evaluation. Results: SEM evaluation showed bone modeling/remodeling between the IAN and implant surface. Fluorochrome area fraction labeling at different times during the healing period showed that bone apposition mainly occurred during the first 2 weeks after implantation. Conclusions: The results obtained showed that bone healing/deposition occurred between the alveolar nerves in contact with a commercially pure titanium implant. No interaction between the nerve and the implant was detected after the 8-week healing period. Appositional bone healing occurred around the nerve bundle structure, restoring the mandibular canal integrity and morphology.

Genotoxic effects of X-rays on keratinized mucosa cells during panoramic dental radiography

Objectives: The aim of this study was to evaluate the genotoxic effects of X-rays on epithelial gingival cells during panoramic dental radiography using a differentiated protocol for the micronucleus test. Methods: 40 healthy individuals who underwent this procedure for diagnostic purposes on request from their dentists agreed to participate in this study. All of them answered a questionnaire before the examination. Epithelial gingival cells were obtained from the keratinized mucosa of the upper dental arcade by gentle scraping with a cervical brush immediately before exposure and 10 days later. Cytological preparations were stained according to the Feulgen-Rossenbeck reaction, counterstained with fast green 1% for 1 min and analysed under a light microscope. Micronuclei, nuclear projections (broken eggs) and degenerative nuclear alterations (pyknosis, karyolysis, karyorrhexis and condensed chromatin) were scored. Results: The frequency of micronuclei was significantly higher after exposure (P < 0.05), as were frequencies of nuclear alterations indicate of apoptosis (P < 0.001). Conclusions: These results indicate that X-ray radiation emitted during panoramic dental radiography induces a genotoxic effect on epithelial gingival cells that increases the frequency of chromosomal damage and nuclear alterations indicative of apoptosis.

Increased Viability of Odontoblast-Like Cells Subjected to Low-Level Laser Irradiation

Studies have shown that the increase of cell metabolism depends on the low level laser therapy (LLLT) parameters used to irradiate the cells. However, the optimal laser dose to up-regulate pulp cell activity remains unknown. Consequently, the aim of this study was to evaluate the metabolic response of odontoblast-like cells (MDPC-23) exposed to different LLLT doses. Cells at 20000 cells/cm(2) were seeded in 24-well plates using plain culture medium (DMEM) and were incubated in a humidified incubator with 5% CO(2) at 37 degrees C. After 24 h, the culture medium was replaced by fresh DMEM supplemented with 5% (stress by nutritional deficit) or 10% fetal bovine serum (FBS). The cells were exposed to different laser doses from a near infrared diode laser prototype designed to provide a uniform irradiation of the wells. The experimental groups were: G1: 1.5 J/cm(2) + 5% FBS; G2: 1.5 J/cm(2) + 10% FBS; G3: 5 J/cm(2) + 5% FBS; G4: 5 J/cm(2) + 10% FBS; G5: 19 J/cm(2) + 5% FBS; G6: 19 J/cm(2) + 10% FBS. LLLT was performed in 3 consecutive irradiation cycles with a 24-hour interval. Non-irradiated cells cultured in DMEM supplemented with either 5 or 10% FBS served as control groups. The analysis of the metabolic response was performed by the MTT assay 3 h after the last irradiation. G1 presented an increase in SDH enzyme activity and differed significantly (Mann-Whitney test, p < 0.05) from the other groups. Analysis by scanning electron microscopy showed normal cell morphology in all groups. Under the tested conditions, LLLT stimulated the metabolic activity of MDPC-23 cultured in DMEM supplemented with 5% FBS and exposed to a laser dose of 1.5 J/cm(2). These findings are relevant for further studies on the action of near infrared lasers on cells with odontoblast phenotype.

Micro Far-Infrared Reflectivity of CaNb(2)O(6) Single Crystal Fibers Grown by the Laser-Heated Pedestal Growth Technique

CaNb(2)O(6) single crystal fibers were grown by the laser-heated pedestal growth technique, directly from the starting reagents. Optically transparent fibers were obtained in the form of rods with elliptical cross-section, free from cracks, impurities, and secondary phases, with an average diameter of 0.4 mm and about 20 mm of length. The fibers grew within the orthorhombic Pbcn columbite structure, with the growth axis nearly parallel to the crystallographic a-direction. The parameters b and c were parallel to the shorter and larger ellipsis axes. A special setup using a microscope was developed to obtain the far-infrared reflectivity spectra of these micrometer-sized fibers, allowing the identification and assignment of 34 of the 38 polar phonons foreseen for the material. From these phonons, the intrinsic dielectric constant ( = 18.2) and quality factor ( of 185 THz) could be estimated, showing the potential of the material for applications in microwave circuitry. These results, along with previous polarized Raman data (Cryst. Growth Des. 2010, 10, 1569), allow us to present a comprehensive set of optical phonon modes and to discuss the potential use of designed CaNb(2)O(6) microcrystals in compact optical devices.

Obtenção de filtro cerâmico a partir da diatomita e casca de arroz carbonizada visando tratamento de efluente têxtil

The effluents released by the textile industry have high concentrations of alkali, carbohydrates, proteins, in addition to colors containing heavy metals. Therefore, a filter was prepared aiming primarily to the removal of color. In order to prepare this filter, rice hulls and diatomite were used, which have in their structure, basically amorphous hydrated silica. The silica exists in three crystalline forms: quartz, tridymite and cristobalite. In accordance with the above considerations, this study was divided into two stages; the first corresponds to the preparation of the filter and the second to carry out the tests in the effluent/filter in order to verify the efficiency of the color removal. First, the raw material was subjected to a chemical analysis and XRD, and then the diatomite was mixed, via humid, with a planetarium windmill with 20 %, 40 %, 60 % and 80 % of rice husk ash. To the mixture, 5 % carboxymethylcellulose (CMC) was added as a binder at room temperature. The samples were uniaxially compacted into metallic matrix of 0.3 x 0.1 cm² of area at a pressure of 167 MPa by means of hydraulic press and then sintered at temperatures of 1,000 °C, 1,200 °C and 1,400 °C for 1 h and submitted to granulometry test using laser, linear retraction, water absorption, apparent porosity and resistance to bending, DTA, TMA and XRD. To examine the pore structure of the samples scanning electron microscope (SEM) was used. Also tests were carried out in a mercury porosimeter to verify the average size of the pores and real density of the samples. In the second stage, samples of the effluent were collected from a local industry, whose name will be preserved, located in Igapó, in the State of Rio Grande do Norte - RN. The effluent was first pretreated before filtration and then subjected to a treatment of flotation. The effluent was then characterized before and after filtration, with parameters of color, turbidity, suspended solids, pH, chemical and biochemical oxygen demand (COD and BOD). Thus, through the XRD analysis the formation of cristobalite α in all samples was observed. The best average size of pore was found to be 1.75 μm with 61.04 % apparent porosity, thus obtaining an average 97.9 % color removal and 99.8 % removal of suspended solid

Al-based anodic oxide films structure observation using field emission gun scanning electron microscopy

In the present work, the anodic oxide films of Al, Al-Cu 4.5% and Al-Si 6.5% alloys are formed using direct and pulse current. In the case of Al-Cu and Al-Si alloys, the electrolyte used contains sulfuric acid and oxalic acid, meanwhile for Al the electrolyte contains sulfuric acid only. Al-Cu alloy was submitted to a heat treatment in order to decrease the effect of inter metallic phase theta upon the anodic film structure. Fractured samples were observed using a field emission gun scanning electron microscope JSM-6330F at (LME)/Brazilian Synchrotron Light Laboratory (LNLS), Campinas, SP, Brazil. The oxide film images enable evaluation of the pore size and form with a resolution similar to the transmission electron microscope (TEM) resolution. It is also observed that the anodizing process using pulse current produces an irregular structure of pore walls, and by direct cur-rent it is produced a rectilinear pore wall. (c) 2005 Elsevier B.V. All rights reserved.

Compósito reforçado com sisal para aplicação na moda

The objective of this research is the fabrication of a composite reinforced with dyed sisal fiber and polyester matrix for application in the fields such as, fashion, clothing, interior textiles; fashion accessories are some of the examples. For the fabrication of the composite, the sisal fibers were subjected to processes such as: chemical treatment with sodium hydroxide (NaOH) in the removal of impurities; bleaching for removing the yellowish color of the natural fiber and dyeing with direct dyes to confer the colors blue, green and orange. The search for new technologies ecologically correct has become a major concern in recent decades. Studies show that composite polymer reinforced by natural fibers is suitable for a large number of applications, and its use is advantageous in terms of economic and ecological. The dyed fibers were cut to a length of 30 mm, is used in the confection of webs. For this purpose, a web preparer by immersion, developed in the Laboratory of Chemical Textile of UFRN. The composite sheets measuring 300 x 300 x3 mm were molded by compression, with unsaturated orthophthalic polyester as matrix, and the samples in sizes 150 x 25 x 3 mm were cut with the aid of a laser machine, to be subjected to traction and flexion. The mechanical properties of traction and flexion in three points were performed in the Laboratory of metal and mechanical tests of Materials Engineering of UFRN. The resulting samples from the tests were evaluated in scanning electron microscope (SEM) at CTGas RN. On the basis of the analysis of the results from the mechanical tests, it was observed that the composite had good mechanical behavior, both in traction as in flexion. Furthermore, it was observed that in the water absorption test, the samples had a different percentage among themselves, this occurred due to the variation of density found in the fibre webs. The images of the SEM showed the failures from the manufacturing process and the adhesion of fibre/matrix. When the samples were prepared with the dyed fibers to be applied in fashion, the results were positive, and it can be concluded that the main objective of this work was achieved

Ultrastructural analysis of the low level laser therapy effects on the lesioned anterior tibial muscle in the Gerbil

Low level laser therapy (LLLT) is known for its positive results but studies on the biological and biomodulator characteristics of the effects produced in the skeletal muscle are Still lacking. In this Study the effects of two laser dosages, 5 or 10 J/cm(2), on the lesioned tibial muscle were compared. Gerbils previously lesioned by 100 g load impact were divided into three groups: GI (n = 5) controls, lesion non-irradiated; GII (n = 5), lesion irradiated with 5 J/Cm(2) and GIII (n = 5), lesion irradiated with 10 J/cm(2), and treated for 7 consecutive days with a laser He-Ne (lambda = 633 rim). After intracardiac perfusion, the muscles were dissected and reduced to small fragments, post-fixed in 1% osmium tetroxide, dehydrated in increasing alcohol concentrations, treated with propylene oxide and embedded in Spurr resin at 60 degrees C. Ultrafine Cuts examined on a transmission electron microscope (Jeol 1010) revealed in the control GI group a large number of altered Muscle fibers with degenerating mitochondria, intercellular substance containing degenerating cell fragments and budding blood capillaries with Underdeveloped endothelial cells. However, groups GII and GIII showed muscle fibers with few altered myofibrils, regularly contoured mitochondria, ample intermembrane spaces and dilated mitochondrial crests. The clean intercellular Substance showed numerous collagen fibers and capillaries with multiple abluminal processes, intraluminal protrusions and several pinocytic vesicles in endothelial cells. it was concluded that laser dosages of 5 or 10 J/cm(2) delivered by laser He-Ne (lambda = 633 rim) during 7 consecutive days increase mitochondrial activity in muscular fibers, activate fibroblasts and macrophages and stimulate angiogenesis, thus suggesting effectivity of laser therapy tinder these experimental conditions. (C) 2009 Elsevier Ltd. All rights reserved.

Low voltage and variable-pressure scanning electron microscopy of fractured composites

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

MRT letter: Extended depth from focus reconstruction method for stretch zone measurement in 15-5PH steel

The stretch zone width (SZW) data for 15-5PH steel CTOD specimens fractured at -150 degrees C to + 23 degrees C temperature were measured based on focused images and 3D maps obtained by extended depth-of-field reconstruction from light microscopy (LM) image stacks. This LM-based method, with a larger lateral resolution, seems to be as effective for quantitative analysis of SZW as scanning electron microscopy (SEM) or confocal scanning laser microscopy (CSLM), permitting to clearly identify stretch zone boundaries. Despite the worst sharpness of focused images, a robust linear correlation was established to fracture toughness (KC) and SZW data for the 15-5PH steel tested specimens, measured at their center region. The method is an alternative to evaluate the boundaries of stretched zones, at a lower cost of implementation and training, since topographic data from elevation maps can be associated with reconstructed image, which summarizes the original contrast and brightness information. Finally, the extended depth-of-field method is presented here as a valuable tool for failure analysis, as a cheaper alternative to investigate rough surfaces or fracture, compared to scanning electron or confocal light microscopes. Microsc. Res. Tech. 75:11551158, 2012. (C) 2012 Wiley Periodicals, Inc.

Analyse titanium surface irradiated with laser, with and without deposited of durapatite

OBJETIVO: Estudar a superfície de implantes osseointegráveis utilizando discos de titânio irradiados com feixe de laser. MÉTODOS: A amostra foi irradiada com feixes de laser de alta intensidade (Nd-YAG), posteriormente depositado hidróxiapatita e submetido a tratamento térmico. Foi analisada sob MEV (Microscópio Eletrônico de Varredura) e realizada análise morfológica qualitativa com microfotografias em vários aumentos. RESULTADOS: A superfície irradiada com laser apresentou deformidade superficial e característica isomórfica; a aplicação de hidroxiapatita pelo método de biomimético aumentou quantitativamente a área da superfície de titânio. CONCLUSÃO: A deposição de hidroxiapatita apresentou melhor característica isomórfica e aumento quantitativo da área superficial estudada, a amostra demonstrou características não encontradas nos implantes disposto no mercado.