Search results for: two-photon fluorescence.

Authors: Lijiang Wang, Wei Wang, Yuhong Xu

Abstract:

Lanthanide-doped upconversion nanoparticles which can convert near-infrared lights to visible lights have attracted growing interest because of their great potentials in fluorescence imaging. Upconversion fluorescence imaging technique with excitation in the near-infrared (NIR) region has been used for imaging of biological cells and tissues. However, improving the detection sensitivity and decreasing the absorption and scattering in biological tissues are as yet unresolved problems. In this present study, a novel NIR-reflected multispectral imaging system was developed for upconversion fluorescent imaging in small animals. Based on this system, we have obtained the high contrast images without the autofluorescence when biocompatible UCPs were injected near the body surface or deeply into the tissue. Furthermore, we have extracted respective spectra of the upconversion fluorescence and relatively quantify the fluorescence intensity with the multispectral analysis. To our knowledge, this is the first time to analyze and quantify the upconversion fluorescence in the small animal imaging.

Keywords: Multispectral imaging, near-infrared, upconversion fluorescence imaging, upconversion nanoparticles.

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Authors: E. Khaleghi, K. Arzani, N. Moallemi, M. Barzegar

Abstract:

This study was conducted to determine effect of water stress on chlorophyll content and chlorophyll fluorescence parameter in young `Dezful- olive trees. Three irrigation regimes (40% ETcrop, 65% ETcrop and 100% ETcrop) were used. After irrigation treatments were applied, some of biochemical parameters including chlorophyll a, b, total chlorophyll, chlorophyll fluorescence and also chlorophyll content index (C.C.I) were measured. Results of Analysis of variance showed that irrigation treatments had significant effect on chlorophylla, total chlorophyll (chl a+b), C.C.I and Fv/Fm ratio. The amount of decreased chlorophyll a and total chlorophyll in plants were received 40% ETcrop were 51.55% and 46.86%, respectively, compared with 100% ETcrop.

Keywords: Evatarnspiration (ETcrop), Chlorophyll Content, Chlorophyll Fluorescence, Water stress, Olive

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Authors: Sanaz Seraj, Shohre Rouhani

Abstract:

Recently, graphene has gained much attention because of its unique optical, mechanical, electrical, and thermal properties. Graphene has been used as a key material in the technological applications in various areas such as sensors, drug delivery, super capacitors, transparent conductor, and solar cell. It has a superior quenching efficiency for various fluorophores. Based on these unique properties, the optical sensors with graphene materials as the energy acceptors have demonstrated great success in recent years. During quenching, the emission of a fluorophore is perturbed by a quencher which can be a substrate or biomolecule, and due to this phenomenon, fluorophore-quencher has been used for selective detection of target molecules. Among fluorescence dyes, 1,8-naphthalimide is well known for its typical intramolecular charge transfer (ICT) and photo-induced charge transfer (PET) fluorophore, strong absorption and emission in the visible region, high photo stability, and large Stokes shift. Derivatives of 1,8-naphthalimides have found applications in some areas, especially fluorescence sensors. Herein, the fluorescence quenching of graphene oxide has been carried out on a naphthalimide dye as a fluorescent probe model. The quenching ability of graphene oxide on naphthalimide dye was studied by UV-VIS and fluorescence spectroscopy. This study showed that graphene is an efficient quencher for fluorescent dyes. Therefore, it can be used as a suitable candidate sensing platform. To the best of our knowledge, studies on the quenching and absorption of naphthalimide dyes by graphene oxide are rare.

Keywords: Fluorescence, graphene oxide, naphthalimide dye, quenching.

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Authors: S. Ashrafpour, T. Tohidi Moghadam, B. Ranjbar

Abstract:

Identifying the nature of protein-nanoparticle interactions and favored binding sites is an important issue in functional characterization of biomolecules and their physiological responses. Herein, interaction of silver nanoparticles with lysozyme as a model protein has been monitored via fluorescence spectroscopy. Formation of complex between the biomolecule and silver nanoparticles (AgNPs) induced a steady state reduction in the fluorescence intensity of protein at different concentrations of nanoparticles. Tryptophan fluorescence quenching spectra suggested that silver nanoparticles act as a foreign quencher, approaching the protein via this residue. Analysis of the Stern-Volmer plot showed quenching constant of 3.73 μM−1. Moreover, a single binding site in lysozyme is suggested to play role during interaction with AgNPs, having low affinity of binding compared to gold nanoparticles. Unfolding studies of lysozyme showed that complex of lysozyme- AgNPs has not undergone structural perturbations compared to the bare protein. Results of this effort will pave the way for utilization of sensitive spectroscopic techniques for rational design of nanobiomaterials in biomedical applications.

Keywords: Nanocarrier, Nanoparticles, Surface Plasmon Resonance, Quenching Fluorescence.

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Authors: P. Soda, G.Iannello

Abstract:

In this paper we address the issue of classifying the fluorescent intensity of a sample in Indirect Immuno-Fluorescence (IIF). Since IIF is a subjective, semi-quantitative test in its very nature, we discuss a strategy to reliably label the image data set by using the diagnoses performed by different physicians. Then, we discuss image pre-processing, feature extraction and selection. Finally, we propose two ANN-based classifiers that can separate intrinsically dubious samples and whose error tolerance can be flexibly set. Measured performance shows error rates less than 1%, which candidates the method to be used in daily medical practice either to perform pre-selection of cases to be examined, or to act as a second reader.

Keywords: Artificial neural networks, computer aided diagnosis, image classification, indirect immuno-fluorescence, pattern recognition.

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Authors: Amanda M. Tadini, Houssam Hajjoul, Gustavo Nicolodelli, Stéphane Mounier, Célia R. Montes, Débora M. B. P. Milori

Abstract:

Soil organic matter (SOM) plays an important role in maintaining soil productivity and accounting for the promotion of biological diversity. The main components of the SOM are the humic substances which can be fractionated according to its solubility in humic acid (HA), fulvic acids (FA) and humin (HU). The determination of the chemical properties of organic matter as well as its interaction with metallic species is an important tool for understanding the structure of the humic fractions. Fluorescence spectroscopy has been studied as a source of information about what is happening at the molecular level in these compounds. Specially, soils of Amazon region are an important ecosystem of the planet. The aim of this study is to understand the molecular and structural composition of HA samples from Spodosol of Amazonia using the fluorescence Emission-Excitation Matrix (EEM) and Time Resolved Fluorescence Spectroscopy (TRFS). The results showed that the samples of HA showed two fluorescent components; one has a more complex structure and the other one has a simpler structure, which was also seen in TRFS through the evaluation of each sample lifetime. Thus, studies of this nature become important because it aims to evaluate the molecular and structural characteristics of the humic fractions in the region that is considered as one of the most important regions in the world, the Amazon.

Keywords: Amazonian soil, characterization, fluorescence, humic acid, lifetime.

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Authors: P. Rattanapoltee, P. Kaewkannetra

Abstract:

According to biodiesel from microalgae is an attractive fuel for several reasons such as renewable, biodegradable and environmental friendly. Thus, this study, green microalgae Scenedesmus acutus PPNK1 isolated from natural water, was selected based on high growth rates, easy cultivation and high lipid content. The Nile red fluorescence method has been successfully applied to the determination of lipids in S. acutus PPNK1. The combination of the method to the lipid composition in algal cells showed the yellow fluorescence under fluorescent microscope. Interestingly, maximum cell numbers and biomass concentration were obtained at 5.44´10⁷ cells/mL and 1.60 g/L when it was cultivated in BG-11 medium while in case of BG-11 with nitrogen deprivation (N 0.25 g/L), accumulated lipid content in cells (44.67%) was achieved that was higher than that found in case of BG-11 medium at about 2 times (22.63%).

Keywords: Biodiesel, Lipid, Microalgae, Nile red.

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Authors: Zhengyu Yan, Yan Yu, Jianqiu Chen

Abstract:

A novel sensing system has been designed for naphthalene detection based on the quenched fluorescence signal of CdS quantum dots. The fluorescence intensity of the system reduced significantly after adding CdS quantum dots to the water pollution model because of the fluorescent static quenching f mechanism. Herein, we have demonstrated the facile methodology can offer a convenient and low analysis cost with the recovery rate as 97.43%-103.2%, which has potential application prospect.

Keywords: CdS quantum dots, modification, detection, naphthalene.

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Authors: R. O. Rominu, C. Sinescu, D.M. Pop, M. Hughes, A. Bradu, M. Rominu, A. Gh. Podoleanu

Abstract:

Bonding has become a routine procedure in several dental specialties – from prosthodontics to conservative dentistry and even orthodontics. In many of these fields it is important to be able to investigate the bonded interfaces to assess their quality. All currently employed investigative methods are invasive, meaning that samples are destroyed in the testing procedure and cannot be used again. We have investigated the interface between human enamel and bonded ceramic brackets non-invasively, introducing a combination of new investigative methods – optical coherence tomography (OCT), fluorescence OCT and confocal microscopy (CM). Brackets were conventionally bonded on conditioned buccal surfaces of teeth. The bonding was assessed using these methods. Three dimensional reconstructions of the detected material defects were developed using manual and semi-automatic segmentation. The results clearly prove that OCT, fluorescence OCT and CM are useful in orthodontic bonding investigations.

Keywords: Optical coherence tomography, Confocal Microscopy, Orthodontic Bonding.

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Authors: Guohua Cao, Xu Dong

Abstract:

X-ray Fluorescence Molecular Imaging (XFMI) holds great promise as a low-cost molecular imaging modality for biomedical applications with high chemical sensitivity. However, for in vivo biomedical applications, a key technical bottleneck is the relatively low chemical sensitivity of XFMI, especially at a reasonably low radiation dose. In laboratory x-ray source based XFMI, one of the main factors that limits the chemical sensitivity of XFMI is the scattered x-rays. We will present our latest findings on improving the chemical sensitivity of XFMI using excitation beam spectrum optimization. XFMI imaging experiments on two mouse-sized phantoms were conducted at three different excitation beam spectra. Our results show that the minimum detectable concentration (MDC) of iodine can be readily increased by five times via excitation spectrum optimization. Findings from this investigation could find use for in vivo pre-clinical small-animal XFMI in the future.

Keywords: Molecular imaging, X-ray fluorescence, chemical sensitivity, X-ray scattering.

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Authors: K. A. Laptinskiy, S. A. Burikov, A. M. Vervald, S. A. Dolenko, T. A. Dolenko

Abstract:

The article presents the results of the application of artificial neural networks to separate the fluorescent contribution of nanodiamonds used as biomarkers, adsorbents and carriers of drugs in biomedicine, from a fluorescent background of own biological fluorophores. The principal possibility of solving this problem is shown. Use of neural network architecture let to detect fluorescence of nanodiamonds against the background autofluorescence of egg white with high accuracy - better than 3 ug/ml.

Keywords: Artificial neural networks, fluorescence, data aggregation.

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Authors: M. A. Elbagermia, A. I. Alajtala, M. Alkerzab

Abstract:

This study compares the quality of different brands of Portland Cement (PC) available in Libyan market. The amounts of chemical constituents like SiO2, Al2O3, Fe2O3, CaO, MgO, SO3 and Lime Saturation Factor (LSF) were determined in accordance with Libyan (L.S.S) and Amrican (A.S.S) Standard Specifications. All the cement studies were found to be good for concrete work especially where no special property is required. The chemical and mineralogical analyses for studied clinker samples show that the dominant phases composition are C3S and C2S while the C3A and C4AF are less abundant.

Keywords: Portland cement, Chemical Composition, Libyan market, X-ray fluorescence.

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Authors: M. Rominu, C. Sinescu, A.G. Podoleanu

Abstract:

The purpose of this study is to present a non invasive method for the marginal adaptation evaluation in class V composite restorations. Standardized class V cavities, prepared in human extracted teeth, were filled with Premise (Kerr) composite. The specimens were thermo cycled. The interfaces were examined by Optical Coherence Tomography method (OCT) combined with the confocal microscopy and fluorescence. The optical configuration uses two single mode directional couplers with a superluminiscent diode as the source at 1300 nm. The scanning procedure is similar to that used in any confocal microscope, where the fast scanning is enface (line rate) and the depth scanning is much slower (at the frame rate). Gaps at the interfaces as well as inside the composite resin materials were identified. OCT has numerous advantages which justify its use in vivo as well as in vitro in comparison with conventional techniques.

Keywords: Class V Cavities, Marginal Adaptation, Optical Coherence Tomography Fluorescence, Confocal Microscopy

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Authors: Chi Man Luk, Ming Kiu Tsang, Chi Fan Chan, Shu Ping Lau

Abstract:

Nitrogen-doped graphene quantum dots (N-GQDs) were fabricated by microwave-assisted hydrothermal technique. The optical properties of the N-GQDs were studied. The luminescence of the N-GQDs can be tuned by varying the excitation wavelength. Furthermore, two-photon luminescence of the N-GQDs excited by near-infrared laser can be obtained. It is shown that N-doping play a key role on two-photon luminescence. The N-GQDs are expected to find application in biological applications including bioimaging and sensing.

Keywords: Graphene quantum dots, nitrogen doping, photoluminescence, two-photon fluorescence.

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Authors: N.E.Galich

Abstract:

We develop new nonlinear methods of immunofluorescence analysis for a sensitive technology of respiratory burst reaction of DNA fluorescence due to oxidative activity in the peripheral blood neutrophils. Histograms in flow cytometry experiments represent a fluorescence flashes frequency as functions of fluorescence intensity. We used the Shannon-Weaver index for definition of neutrophils- biodiversity and Hurst index for definition of fractal-s correlations in immunofluorescence for different donors, as the basic quantitative criteria for medical diagnostics of health status. We analyze frequencies of flashes, information, Shannon entropies and their fractals in immunofluorescence networks due to reduction of histogram range. We found the number of simplest universal correlations for biodiversity, information and Hurst index in diagnostics and classification of pathologies for wide spectra of diseases. In addition is determined the clear criterion of a common immunity and human health status in a form of yes/no answers type. These answers based on peculiarities of information in immunofluorescence networks and biodiversity of neutrophils. Experimental data analysis has shown the existence of homeostasis for information entropy in oxidative activity of DNA in neutrophil nuclei for all donors.

Keywords: blood and cells fluorescence in diagnostics ofdiseases, cytometric histograms, entropy and information in fractalnetworks of oxidative activity of DNA, long-range chromosomalcorrelations in living cells.

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Authors: Blessing Ojeme, Frederick Quinn, Russell Karls, Shannon Quinn

Abstract:

The detection and segmentation of mitochondria from fluorescence microscopy is crucial for understanding the complex structure of the nervous system. However, the constant fission and fusion of mitochondria and image distortion in the background make the task of detection and segmentation challenging. Although there exists a number of open-source software tools and artificial intelligence (AI) methods designed for analyzing mitochondrial images, the availability of only a few combined expertise in the medical field and AI required to utilize these tools poses a challenge to its full adoption and use in clinical settings. Motivated by the advantages of automated methods in terms of good performance, minimum detection time, ease of implementation, and cross-platform compactibility, this study proposes a fully automated framework for the detection and segmentation of mitochondria using both image shape information and descriptive statistics. Using the low-cost, open-source Python and OpenCV library, the algorithms are implemented in three stages: pre-processing; image binarization; and coarse-to-fine segmentation. The proposed model is validated using the fluorescence mitochondrial dataset. Ground truth labels generated using Labkit were also used to evaluate the performance of our detection and segmentation model using precision, recall and rand index. The study produces good detection and segmentation results and reports the challenges encountered during the image analysis of mitochondrial morphology from the fluorescence mitochondrial dataset. A discussion on the methods and future perspectives of fully automated frameworks concludes the paper.

Keywords: 2D, Binarization, CLAHE, detection, fluorescence microscopy, mitochondria, segmentation.

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Authors: M. Abdeshahian, M. Nabipour, M. Meskarbashee

Abstract:

To investigate effect of salt stress on Chlorophyll fluorescence four cultivars (fong,star,chamran and kharchia) of wheat (Triticum aestivum) plants subjected to salinity levels ( control,8,12 and 16 dsm-1 ) from one week after emergence to the end of stem elongation under greenhouse condition . results showed that quantum yield of photosystem II from light adopted leaves (ΦPSII), Photochemical quenching (qP) ,quantum yield of dark adopted leaves (fv/fm) and non photochemical quenching (NPq) were affected by salt stress . Salinity levels affected photosynthetic rate. Star and fong cultivars showed minimum and maximum levels of photosynthetic rate in respectively. Minimum photosynthetic rate differences between levels of salinity were shown in Kharchia. Shoot dry matter of all cultivars decreased by increasing salinity levels. Results showed that non photochemical quenching by salinity levels attribute to the decreases in shoot dry matter.

Keywords: salt stress, wheat, chlorophyll fluorescence, photosynthesis , shoot dry matter .

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Authors: Shigenori Togashi, Kei Takenaka

Abstract:

We developed new processes which can collect and detect rapidly airborne pathogens such as the avian flu virus for the pandemic prevention. The fluorescence antibody technique is known as one of high-sensitive detection methods for viruses, but this needs up to a few hours to bind sufficient fluorescence dyes to viruses for detection. In this paper, we developed a mist-labeling can detect substitution viruses in a short time to improve the binding rate of fluorescent dyes and substitution viruses by the micro reaction process. Moreover, we developed the rapid detection system with the above “mist labeling”. The detection system set with a sampling bag collecting patient’s breath and a cartridge can detect automatically pathogens within 10 minutes.

Keywords: Viruses, Sampler, Mist, Detection, Fluorescent dyes, Microreaction.

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Authors: Fu E. Tang, Daoyi Chen

Abstract:

Experimental investigations were carried out in the Manchester Tidal flow Facility (MTF) to study the flow patterns in the region around and adjacent to a hypothetical headland in tidal (oscillatory) ambient flow. The Planar laser-induced fluorescence (PLIF) technique was used for visualization, with fluorescent dye released at specific points around the headland perimeter and in its adjacent recirculation zone. The flow patterns can be generalized into the acceleration, stable flow and deceleration stages for each halfcycle, with small variations according to location, which are more distinct for low Keulegan-Carpenter number (KC) cases. Flow patterns in the mixing region are unstable and complex, especially in the recirculation zone. The flow patterns are in agreement with previous visualizations, and support previous results in steady ambient flow. It is suggested that the headland lee could be a viable location for siting of pollutant outfalls.

Keywords: Planar laser-induced Fluorescence, recirculation zone, tidal flow, wake flows

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Authors: Alessandro Bevilacqua, Alessandro Gherardi, Filippo Piccinini

Abstract:

Image mosaicing techniques are usually employed to offer researchers a wider field of view of microscopic image of biological samples. a mosaic is commonly achieved using automated microscopes and often with one “color" channel, whether it refers to natural or fluorescent analysis. In this work we present a method to achieve three subsequent mosaics of the same part of a stem cell culture analyzed in phase contrast and in fluorescence, with a common non-automated inverted microscope. The mosaics obtained are then merged together to mark, in the original contrast phase images, nuclei and cytoplasm of the cells referring to a mosaic of the culture, rather than to single images. The experiments carried out prove the effectiveness of our approach with cultures of cells stained with calcein (green/cytoplasm and nuclei) and hoechst (blue/nuclei) probes.

Keywords: Microscopy, image mosaicing, fluorescence, stem cells.

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Authors: S.F.S. Draman, R. Daik, A. Musa

Abstract:

Three sulphonic acid-doped polyanilines were synthesized through chemical oxidation at low temperature (0-5 oC) and potential of these polymers as sensing agent for O2 gas detection in terms of fluorescence quenching was studied. Sulphuric acid, dodecylbenzene sulphonic acid (DBSA) and camphor sulphonic acid (CSA) were used as doping agents. All polymers obtained were dark green powder. Polymers obtained were characterized by Fourier transform infrared spectroscopy, ultraviolet-visible absorption spectroscopy, thermogravimetry analysis, elemental analysis, differential scanning calorimeter and gel permeation chromatography. Characterizations carried out showed that polymers were successfully synthesized with mass recovery for sulphuric aciddoped polyaniline (SPAN), DBSA-doped polyaniline (DBSA-doped PANI) and CSA-doped polyaniline (CSA-doped PANI) of 71.40%, 75.00% and 39.96%, respectively. Doping level of SPAN, DBSAdoped PANI and CSA-doped PANI were 32.86%, 33.13% and 53.96%, respectively as determined based on elemental analysis. Sensing test was carried out on polymer sample in the form of solution and film by using fluorescence spectrophotometer. Samples of polymer solution and polymer film showed positive response towards O2 exposure. All polymer solutions and films were fully regenerated by using N2 gas within 1 hour period. Photostability study showed that all samples of polymer solutions and films were stable towards light when continuously exposed to xenon lamp for 9 hours. The relative standard deviation (RSD) values for SPAN solution, DBSA-doped PANI solution and CSA-doped PANI solution for repeatability were 0.23%, 0.64% and 0.76%, respectively. Meanwhile RSD values for reproducibility were 2.36%, 6.98% and 1.27%, respectively. Results for SPAN film, DBSAdoped PANI film and CSA-doped PANI film showed the same pattern with RSD values for repeatability of 0.52%, 4.05% and 0.90%, respectively. Meanwhile RSD values for reproducibility were 2.91%, 10.05% and 7.42%, respectively. The study on effect of the flow rate on response time was carried out using 3 different rates which were 0.25 mL/s, 1.00 mL/s and 2.00 mL/s. Results obtained showed that the higher the flow rate, the shorter the response time.

Keywords: conjugated polymer, doping, fluorescence quenching, oxygen gas.

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Authors: Ndeye Rokhaya Faye, Ibtissem Gammoudi, Fabien Moroté, Christine Grauby-Heywang, TouriaCohen-Bouhacina

Abstract:

In this study we investigate silica nanoparticle (SiO2- NP) effects on the structure and phase properties of supported lipid monolayers and bilayers, coupling surface pressure measurements, fluorescence microscopy and atomic force microscopy. SiO2-NPs typically in size range of 10nm to 100 nm in diameter are tested. Our results suggest first that lipid molecules organization depends to their nature. Secondly, lipid molecules in the vinicity of big aggregates nanoparticles organize in liquid condensed phase whereas small aggregates are localized in both fluid liquid-expanded (LE) and liquid-condenced (LC). We demonstrated also by atomic force microscopy that by measuring friction forces it is possible to get information as if nanoparticle aggregates are recovered or not by lipid monolayers and bilayers.

Keywords: Atomic force microscopy, fluorescence microscopy, Langmuir films, silica nanoparticles, supported membrane models.

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Authors: Bireswar Paul, Amitava Datta

Abstract:

Indoor air environment is a big concern in the last few decades in the developing countries, with increased focus on monitoring the air quality. In this work, an experimental study has been conducted to establish the existence of carbon nanoparticles below the size range of 10 nm in the non-sooting zone of a LPG/air partially premixed flame. Mainly, four optical techniques, UV absorption spectroscopy, fluorescence spectroscopy, dynamic light scattering and TEM have been used to characterize and measure the size of carbon nanoparticles in the sampled materials collected from the inner surface of the flame front. The existence of the carbon nanoparticles in the sampled material has been confirmed with the typical nature of the absorption and fluorescence spectra already reported in the literature. The band gap energy shows that the particles are made up of three to six aromatic rings. The size measurement by DLS technique also shows that the particles below the size range of 10 nm. The results of DLS are also corroborated by the TEM image of the same material. 

Keywords: Indoor air, carbon nanoparticles, LPG, partially premixed flame, optical techniques.

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Authors: Yunxia Zhang, Jiti Zhou, Jianbo Guo, Xiuhong Zhang, Lihong Zhao, Shouzhi Yuan

Abstract:

Novel nitrogen removal technologies via nitrite pathway attract increasing interest in recent years. In this study, batch experiments were performed to investigate nitrite accumulation characteristics and shifts in nitrifying community structure at different growth environments including ammonia concentration, pH and alkalinity. It was found that nitrite accumulation ratios were maintained at around 95% at studied conditions, and the optimum pH and Alk/N (ratio between alkalinity and nitrogen) for ammonium oxidization were 8.5 and 8.33, respectively. Fluorescence in situ hybridization analysis of nitrifying bacteria showed that high free ammonia (from influent ammonium or caused by high pH) significantly altered the structure of nitrifying community, leading to abundance of ammonia-oxidizing bacteria (AOB), especially Nitrososmonas, and inhibition of nitrite-oxidizing bacteria (NOB). The results suggest that free ammonia plays more important role than other studied conditions on nitrite accumulation.

Keywords: Partial nitrification, Nitrite accumulation, Nitrifyingbacteria, Fluorescence in situ hybridization (FISH).

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Authors: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

Abstract:

Celiac disease (CD) is an immune system disorder that is related to eating gluten. As gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over a 8-week long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories; giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively, when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first 4 weeks and then decays to 63%.

Keywords: Aptasensor, PEG, rGO, FAM, RM, ELISA.

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Authors: Mohammad Emran, Colton Wayne, Shannon M Koehler, P. Stephen Almond, Haroon Patel

Abstract:

Purpose: Assessment of anastomotic perfusion in Hirschsprung disease using Indocyanine Green (ICG)-near-infrared (NIR) fluorescence angiography. Introduction: Anastomotic stricture and leak are well-known complications of Hirschsprung pull-through procedures. Complications are due to tension, infection, and/or poor perfusion. While a surgeon can visually determine and control the amount of tension and contamination, assessment of perfusion is subject to surgeon determination. Intraoperative use of ICG-NIR enhances this decision-making process by illustrating perfusion intensity and adequacy in the pulled-through bowel segment. This technique, proven to reduce anastomotic stricture and leak in adults, has not been studied in children to our knowledge. ICG, an FDA approved, nontoxic, non-immunogenic, intravascular (IV) dye, has been used in adults and children for over 60 years, with few side effects. ICG-NIR was used in this report to demonstrate the adequacy of perfusion during transanal pullthrough for Hirschsprung’s disease. Method: 8 patients with Hirschsprung disease were evaluated with ICG-NIR technology. Levels of affected area ranged from sigmoid to total colonic Hirschsprung disease. After leveling, but prior to anastomosis, ICG was administered at 1.25 mg (< 2 mg/kg) and perfusion visualized using an NIR camera, before and during anastomosis. Video and photo imaging was performed and perfusion of the bowel was compared to surrounding tissues. This showed the degree of perfusion and demarcation of perfused and non-perfused bowel. The anastomosis was completed uneventfully and the patients all did well. Results: There were no complications of stricture or leak. 5 of 8 patients (62.5%) had modification of the plan based on ICG-NIR imaging. Conclusion: Technologies that enhance surgeons’ ability to visualize bowel perfusion prior to anastomosis in Hirschsprung’s patients may help reduce post-operative complications. Further studies are needed to assess the potential benefits.

Keywords: Colonic anastomosis, fluorescence angiography, Hirschsprung disease, pediatric surgery, SPY, ICG, NIR.

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Authors: C. Sinescu, M. Negrutiu, M. Romînu, C. Haiduc, E. Petrescu, M. Leretter, A.G. Podoleanu

Abstract:

Optical Coherence Tomography (OCT) combined with the Confocal Microscopy, as a noninvasive method, permits the determinations of materials defects in the ceramic layers depth. For this study 256 anterior and posterior metal and integral ceramic fixed partial dentures were used, made with Empress (Ivoclar), Wollceram and CAD/CAM (Wieland) technology. For each investigate area 350 slices were obtain and a 3D reconstruction was perform from each stuck. The Optical Coherent Tomography, as a noninvasive method, can be used as a control technique in integral ceramic technology, before placing those fixed partial dentures in the oral cavity. The purpose of this study is to evaluate the capability of En face Optical Coherence Tomography (OCT) combined with a fluorescent method in detection and analysis of possible material defects in metalceramic and integral ceramic fixed partial dentures. As a conclusion, it is important to have a non invasive method to investigate fixed partial prostheses before their insertion in the oral cavity in order to satisfy the high stress requirements and the esthetic function.

Keywords: Ceramic Fixed Partial Dentures, Material Defects, En face Optical Coherence Tomography, Fluorescence.

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Authors: R. A. Shahmiri, O. C. Standard, J. N. Hart, C. C. Sorrell

Abstract:

The ceramic yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) has been used as a dental biomaterial for several decades. The strength and toughness of this material can be accounted for by its toughening mechanisms, which include transformation toughening, crack deflection, zone shielding, contact shielding, and crack bridging. Prevention of crack propagation is of critical importance in high-fatigue situations, such as those encountered in mastication and para-function. However, the poor translucence of Y-TZP in polycrystalline form is such that it may not meet the aesthetic requirements due to its white/grey appearance. To improve the optical properties of Y-TZP, more detailed study of the optical properties is required; in particular, precise evaluation of the refractive index, absorption coefficient, and scattering coefficient are necessary. The measurement of the optical parameters has been based on the assumption that light scattered from biological media is isotropically distributed over all angles. In fact, the optical behavior of real biological materials depends on the angular scattering of light due to the anisotropic nature of the materials. The purpose of the present work is to evaluate the optical properties (including color, opacity/translucence, scattering, and fluorescence) of zirconia dental ceramics and their control through modification of the chemical composition, phase composition, and surface microstructure.

Keywords: Optical properties, opacity/translucence, scattering, fluorescence, chemical composition, phase composition, surface microstructure.

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

Abstract:

Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: Image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization.

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Authors: Kheireddine El-Boubbou

Abstract:

Drug delivery to target human acute myeloid leukemia (AML) using a nanoparticulate chemotherapeutic formulation that can deliver drugs selectively to AML cancer is hugely needed. In this work, we report the development of a nanoformulation made of polymeric-stabilized multifunctional magnetic iron oxide nanoparticles (PMNP) loaded with the anticancer drug Doxorubicin (Dox) as a promising drug carrier to treat AML. Dox@PMNP conjugates simultaneously exhibited high drug content, maximized fluorescence, and excellent release properties. Nanoparticulate uptake and cell death following addition of Dox@PMNPs were then evaluated in different types of human AML target cells, as well as on normal human cells. While the unloaded MNPs were not toxic to any of the cells, Dox@PMNPs were found to be highly toxic to the different AML cell lines, albeit at different inhibitory concentrations (IC₅₀ values), but showed very little toxicity towards the normal cells. In comparison, free Dox showed significant potency concurrently to all the cell lines, suggesting huge potentials for the use of Dox@PMNPs as selective AML anticancer cargos. Live confocal imaging, fluorescence and electron microscopy confirmed that Dox is indeed delivered to the nucleus in relatively short periods of time, causing apoptotic cell death. Importantly, this targeted payload may potentially enhance the effectiveness of the drug in AML patients and may further allow physicians to image leukemic cells exposed to Dox@PMNPs using MRI.

Keywords: Magnetic nanoparticles, drug delivery, acute myeloid leukemia, iron oxide, cancer nanotherapy.

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