Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30

Fluorescence Related Abstracts

30 Using Artificial Neural Networks for Optical Imaging of Fluorescent Biomarkers

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


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, Biomarkers, data aggregation, Fluorescence

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29 Determination of the Botanical Origin of Honey by the Artificial Neural Network Processing of PARAFAC Scores of Fluorescence Data

Authors: Lea Lenhardt, Ivana Zeković, Tatjana Dramićanin, Miroslav D. Dramićanin


Fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC) and artificial neural networks (ANN) were used for characterization and classification of honey. Excitation emission spectra were obtained for 95 honey samples of different botanical origin (acacia, sunflower, linden, meadow, and fake honey) by recording emission from 270 to 640 nm with excitation in the range of 240-500 nm. Fluorescence spectra were described with a six-component PARAFAC model, and PARAFAC scores were further processed with two types of ANN’s (feed-forward network and self-organizing maps) to obtain algorithms for classification of honey on the basis of their botanical origin. Both ANN’s detected fake honey samples with 100% sensitivity and specificity.

Keywords: Artificial Neural Networks, Fluorescence, honey, PARAFAC

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28 Analisys of Cereal Flours by Fluorescence Spectroscopy and PARAFAC

Authors: Lea Lenhardt, Ivana Zeković, Tatjana Dramićanin, Miroslav D. Dramićanin


Rapid and sensitive analytical technologies for food analysis are needed to respond to the growing public interest in food quality and safety. In this context, fluorescence spectroscopy offers several inherent advantages for the characterization of food products: high sensitivity, low price, objective, relatively fast and non-destructive. The objective of this work was to investigate the potential of fluorescence spectroscopy coupled with multi-way technique for characterization of cereal flours. Fluorescence landscape also known as excitation-emission matrix (EEM) spectroscopy utilizes multiple-color illumination, with the full fluorescence spectrum recorded for each excitation wavelength. EEM was measured on various types of cereal flours (wheat, oat, barley, rye, corn, buckwheat and rice). Obtained spectra were analyzed using PARAllel FACtor analysis (PARAFAC) in order to decompose the spectra and identify underlying fluorescent components. Results of the analysis indicated the presence of four fluorophores in cereal flours. It has been observed that relative concentration of fluorophores varies between different groups of flours. Based on these findings we can conclude that application of PARAFAC analysis on fluorescence data is a good foundation for further qualitative analysis of cereal flours.

Keywords: Fluorescence, PARAFAC, cereals, fluors

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27 An Activatable Prodrug for the Treatment of Metastatic Tumors

Authors: Hyunseung Lee, Eun-Joong Kim, Sankarprasad Bhuniya, Hyun Min Kim, Chaejoon Cheong, Su-khendu Maiti, Kwan Soo Hong, Jong Seung Kim


Metastatic cancers have historically been difficult to treat. However, metastatic tumors have been found to have high levels of reactive oxygen species such as hydrogen peroxide (H2O2), supporting the hypothesis that a prodrug could be activated by intracellular H2O2 and lead to a potential anti-metastatic therapy. In this study, prodrug 7 was designed to be activated by H2O2-mediated boronate oxidation, resulting in activation of the fluorophore for detection and release of the therapeutic agent, SN-38. Drug release from prodrug 7 was investigated by monitoring fluorescence after addition of H2O2 to the cancer cells. Prodrug 7 activated by H2O2 selectively inhibited tumor cell growth. Furthermore, intratracheally administered prodrug 7 showed effective anti-tumor activity in a mouse model of metastatic lung disease. Thus, this H2O2-responsive prodrug has therapeutic potential as a novel treatment for metastatic cancer via cellular imaging with fluorescence as well as selective release of the anti-cancer drug, SN-38.

Keywords: Fluorescence, Hydrogen Peroxide, prodrug, metastatic tumors

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26 An Activatable Theranostic for Targeted Cancer Therapy and Imaging

Authors: Hyunseung Lee, Eun-Joong Kim, Sankarprasad Bhuniya, Kwan Soo Hong, Jong Seung Kim, Sukhendu Maiti, Jonathan L. Sessler


A new theranostic strategy is described. It is based on the use of an “all in one” prodrug, namely the biotinylated piperazine-rhodol conjugate 4a. This conjugate, which incorporates the anticancer drug SN-38, undergoes self-immolative cleavage when exposed to biological thiols. This leads to the tumor-targeted release of the active SN-38 payload along with fluorophore 1a. This release is made selective as the result of the biotin functionality. Fluorophore 1a is 32-fold more fluorescent than prodrug 4a. It permits the delivery and release of the SN-38 payload to be monitored easily in vitro and in vivo, as inferred from cell studies and ex vivo analyses of mice xenografts derived HeLa cells, respectively. Prodrug 4a also displays anticancer activity in the HeLa cell murine xenograft tumor model. On the basis of these findings we suggest that the present strategy, which combines within a single agent the key functions of targeting, release, imaging, and treatment, may have a role to play in cancer diagnosis and therapy.

Keywords: Cancer Therapy, Fluorescence, Theranostic, prodrug

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25 Application of Flow Cytometry for Detection of Influence of Abiotic Stress on Plants

Authors: Dace Grauda, Inta Belogrudova, Isaak Rashal, Alexei Katashev, Linda Lancere


The goal of study was the elaboration of easy applicable flow cytometry method for detection of influence of abiotic stress factors on plants, which could be useful for detection of environmental stresses in urban areas. The lime tree Tillia vulgaris H. is a popular tree species used for urban landscaping in Europe and is one of the main species of street greenery in Riga, Latvia. Tree decline and low vitality has observed in the central part of Riga. For this reason lime trees were select as a model object for the investigation. During the period of end of June and beginning of July 12 samples from different urban environment locations as well as plant material from a greenhouse were collected. BD FACSJazz® cell sorter (BD Biosciences, USA) with flow cytometer function was used to test viability of plant cells. The method was based on changes of relative fluorescence intensity of cells in blue laser (488 nm) after influence of stress factors. SpheroTM rainbow calibration particles (3.0–3.4 μm, BD Biosciences, USA) in phosphate buffered saline (PBS) were used for calibration of flow cytometer. BD PharmingenTM PBS (BD Biosciences, USA) was used for flow cytometry assays. The mean fluorescence intensity information from the purified cell suspension samples was recorded. Preliminary, multiple gate sizes and shapes were tested to find one with the lowest CV. It was found that low CV can be obtained if only the densest part of plant cells forward scatter/side scatter profile is analysed because in this case plant cells are most similar in size and shape. The young pollen cells in one nucleus stage were found as the best for detection of influence of abiotic stress. For experiments only fresh plant material was used– the buds of Tillia vulgaris with diameter 2 mm. For the cell suspension (in vitro culture) establishment modified protocol of microspore culture was applied. The cells were suspended in the MS (Murashige and Skoog) medium. For imitation of dust of urban area SiO2 nanoparticles with concentration 0.001 g/ml were dissolved in distilled water. Into 10 ml of cell suspension 1 ml of SiO2 nanoparticles suspension was added, then cells were incubated in speed shaking regime for 1 and 3 hours. As a stress factor the irradiation of cells for 20 min by UV was used (Hamamatsu light source L9566-02A, L10852 lamp, A10014-50-0110), maximum relative intensity (100%) at 365 nm and at ~310 nm (75%). Before UV irradiation the suspension of cells were placed onto a thin layer on a filter paper disk (diameter 45 mm) in a Petri dish with solid MS media. Cells without treatment were used as a control. Experiments were performed at room temperature (23-25 °C). Using flow cytometer BS FACS Software cells plot was created to determine the densest part, which was later gated using oval-shaped gate. Gate included from 95 to 99% of all cells. To determine relative fluorescence of cells logarithmic fluorescence scale in arbitrary fluorescence units were used. 3x103 gated cells were analysed from the each sample. The significant differences were found among relative fluorescence of cells from different trees after treatment with SiO2 nanoparticles and UV irradiation in comparison with the control.

Keywords: Fluorescence, flow cytometry, UV irradiation, SiO2 nanoparticles

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24 Encoded Fiber Optic Sensors for Simultaneous Multipoint Sensing

Authors: C. Babu Rao, Pandian Chelliah


Owing to their reliability, a number of fluorescent spectra based fiber optic sensors have been developed for detection and identification of hazardous chemicals such as explosives, narcotics etc. In High security regions, such as airports, it is important to monitor simultaneously multiple locations. This calls for deployment of a portable sensor at each location. However, the selectivity and sensitivity of these techniques depends on the spectral resolution of the spectral analyzer. The better the resolution the larger the repertoire of chemicals that can be detected. A portable unit will have limitations in meeting these requirements. Optical fibers can be employed for collecting and transmitting spectral signal from the portable sensor head to a sensitive central spectral analyzer (CSA). For multipoint sensing, optical multiplexing of multiple sensor heads with CSA has to be adopted. However with multiplexing, when one sensor head is connected to CSA, the rest may remain unconnected for the turn-around period. The larger the number of sensor heads the larger this turn-around time will be. To circumvent this imitation, we propose in this paper, an optical encoding methodology to use multiple portable sensor heads connected to a single CSA. Each portable sensor head is assigned an unique address. Spectra of every chemical detected through this sensor head, are encoded by its unique address and can be identified at the CSA end. The methodology proposed is demonstrated through a simulation using Matlab SIMULINK.

Keywords: Fluorescence, optical encoding, multipoint sensing

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23 Online Monitoring of Airborne Bioaerosols Released from a Composting, Green Waste Site

Authors: John Sodeau, David O'Connor, Shane Daly, Stig Hellebust


This study is the first to employ the online WIBS (Waveband Integrated Biosensor Sensor) technique for the monitoring of bioaerosol emissions and non-fluorescing “dust” released from a composting/green waste site. The purpose of the research was to provide a “proof of principle” for using WIBS to monitor such a location continually over days and nights in order to construct comparative “bioaerosol site profiles”. Current impaction/culturing methods take many days to achieve results available by the WIBS technique in seconds.The real-time data obtained was then used to assess variations of the bioaerosol counts as a function of size, “shape”, site location, working activity levels, time of day, relative humidity, wind speeds and wind directions. Three short campaigns were undertaken, one classified as a “light” workload period, another as a “heavy” workload period and finally a weekend when the site was closed. One main bioaerosol size regime was found to predominate: 0.5 micron to 3 micron with morphologies ranging from elongated to elipsoidal/spherical. The real-time number-concentration data were consistent with an Andersen sampling protocol that was employed at the site. The number-concentrations of fluorescent particles as a proportion of total particles counted amounted, on average, to ~1% for the “light” workday period, ~7% for the “heavy” workday period and ~18% for the weekend. The bioaerosol release profiles at the weekend were considerably different from those monitored during the working weekdays.

Keywords: Bioaerosols, Composting, Fluorescence, particle counting in real-time

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22 Photophysical Study of Pyrene Butyric Acid in Aqueous Ionic Liquid

Authors: Pratap K. Chhotaray, Jitendriya Swain, Ashok Mishra, Ramesh L. Gardas


Ionic liquids (ILs) are molten salts, consist predominantly of ions and found to be liquid below 100°C. The unparalleled growing interest in ILs is based upon their never ending design flexibility. The use of ILs as a co-solvent in binary as well as a ternary mixture with molecular solvents multifold it’s utility. Since polarity is one of the most widely applied solvent concepts which represents simple and straightforward means for characterizing and ranking the solvent media, its study for a binary mixture of ILs is crucial for its widespread application and development. The primary approach to the assessment of solution phase intermolecular interactions, which generally occurs on the picosecond to nanosecond time scales, is to exploit the optical response of photophysical probe. Pyrene butyric acid (PBA) is used as fluorescence probe due to its high quantum yield, longer lifetime and high solvent polarity dependence of fluorescence spectra. Propylammonium formate (PAF) is the IL used for this study. Both the UV-absorbance spectra and steady state fluorescence intensity study of PBA in different concentration of aqueous PAF, reveals that with an increase in PAF concentration, both the absorbance and fluorescence intensity increases which indicate the progressive solubilisation of PBA. Whereas, near about 50% of IL concentration, all of the PBA molecules get solubilised as there are no changes in the absorbance and fluorescence intensity. Furthermore, the ratio II/IV, where the band II corresponds to the transition from S1 (ν = 0) to S0 (ν = 0), and the band IV corresponds to transition from S1 (ν = 0) to S0 (ν = 2) of PBA, indicates that the addition of water into PAF increases the polarity of the medium. Time domain lifetime study shows an increase in lifetime of PBA towards the higher concentration of PAF. It can be attributed to the decrease in non-radiative rate constant at higher PAF concentration as the viscosity is higher. The monoexponential decay suggests that homogeneity of solvation environment whereas the uneven width at full width at half maximum (FWHM) indicates there might exist some heterogeneity around the fluorophores even in the water-IL mixed solvents.

Keywords: Fluorescence, polarity, ionic liquid, lifetime, pyrene butyric acid

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21 Advanced Real-Time Fluorescence Imaging System for Rat's Femoral Vein Thrombosis Monitoring

Authors: Chul Gyu Song, Sang Hun Park


Artery and vein occlusion changes observed in patients and experimental animals are unexplainable symptoms. As the fat accumulated in cardiovascular ruptures, it causes vascular blocking. Likewise, early detection of cardiovascular disease can be useful for treatment. In this study, we used the mouse femoral occlusion model to observe the arterial and venous occlusion changes without darkroom. We observed the femoral arterial flow pattern changes by proposed fluorescent imaging system using an animal model of thrombosis. We adjusted the near-infrared light source current in order to control the intensity of the fluorescent substance light. We got the clear fluorescent images and femoral artery flow pattern were measured by a 5-minute interval. The result showed that the fluorescent substance flowing in the femoral arteries were accumulated in thrombus as time passed, and the fluorescence of other vessels gradually decreased.

Keywords: Fluorescence, thrombus, arteries, femoral

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20 A Fluorescent Polymeric Boron Sensor

Authors: Soner Cubuk, Mirgul Kosif, M. Vezir Kahraman, Ece Kok Yetimoglu


Boron is an essential trace element for the completion of the life circle for organisms. Suitable methods for the determination of boron have been proposed, including acid - base titrimetric, inductively coupled plasma emission spectroscopy flame atomic absorption and spectrophotometric. However, the above methods have some disadvantages such as long analysis times, requirement of corrosive media such as concentrated sulphuric acid and multi-step sample preparation requirements and time-consuming procedures. In this study, a selective and reusable fluorescent sensor for boron based on glycosyloxyethyl methacrylate was prepared by photopolymerization. The response characteristics such as response time, pH, linear range, limit of detection were systematically investigated. The excitation/emission maxima of the membrane were at 378/423 nm, respectively. The approximate response time was measured as 50 sec. In addition, sensor had a very low limit of detection which was 0.3 ppb. The sensor was successfully used for the determination of boron in water samples with satisfactory results.

Keywords: Fluorescence, Boron, Photopolymerization, polymeric sensor

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19 Fluorescence in situ Hybridization (FISH) Detection of Bacteria and Archaea in Fecal Samples

Authors: Maria Nejjari, Michel Cloutier, Guylaine Talbot, Martin Lanthier


The fluorescence in situ hybridization (FISH) is a staining technique that allows the identification, detection and quantification of microorganisms without prior cultivation by means of epifluorescence and confocal laser scanning microscopy (CLSM). Oligonucleotide probes have been used to detect bacteria and archaea that colonize the cattle and swine digestive systems. These bacterial strains have been obtained from fecal samples issued from cattle manure and swine slurry. The collection of these samples has been done at 3 different pit’s levels A, B and C with same height. Two collection depth levels have been taken in consideration, one collection level just under the pit’s surface and the second one at the bottom of the pit. Cells were fixed and FISH was performed using oligonucleotides of 15 to 25 nucleotides of length associated with a fluorescent molecule Cy3 or Cy5. The double hybridization using Cy3 probe targeting bacteria (Cy3-EUB338-I) along with a Cy5 probe targeting Archaea (Gy5-ARCH915) gave a better signal. The CLSM images show that there are more bacteria than archaea in swine slurry. However, the choice of fluorescent probes is critical for getting the double hybridization and a unique signature for each microorganism. FISH technique is an easy way to detect pathogens like E. coli O157, Listeria, Salmonella that easily contaminate water streams, agricultural soils and, consequently, food products and endanger human health.

Keywords: Fish, Bacteria, Archaea, Detection, Fluorescence

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18 A Turn-on Fluorescent Sensor for Pb(II)

Authors: Soner Cubuk, M. Vezir Kahraman, Ece Kok Yetimoglu, Neşe Taşci


Lead(II) is one of the most toxic environmental pollutants in the world, due to its high toxicity and non-biodegradability. Lead exposure causes severe risks to human health such as central brain damages, convulsions, kidney damages, and even death. To determine lead(II) in environmental or biological samples, scientists use atomic absorption spectrometry (AAS), inductively coupled plasma mass spectrometry (ICPMS), fluorescence spectrometry and electrochemical techniques. Among these systems the fluorescence spectrometry and fluorescent chemical sensors have attracted considerable attention because of their good selectivity and high sensitivity. The fluorescent polymers usually contain covalently bonded fluorophores. In this study imidazole based UV cured polymeric film was prepared and designed to act as a fluorescence chemo sensor for lead (II) analysis. The optimum conditions such as influence of pH value and time on the fluorescence intensity of the sensor have also been investigated. The sensor was highly sensitive with a detection limit as low as 1.87 × 10−8 mol L-1 and it was successful in the determination of Pb(II) in water samples.

Keywords: Fluorescence, Photopolymerization, polymeric sensor, lead(II)

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17 Development of Polymeric Fluorescence Sensor for the Determination of Bisphenol-A

Authors: Soner Cubuk, M. Vezir Kahraman, Ece Kok Yetimoglu, Neşe Taşci


Bisphenol-A (BPA), 2,2-bis(4-hydroxyphenly)propane, is one of the highest usage volume chemicals in the world. Studies showed that BPA maybe has negative effects on the central nervous system, immune and endocrine systems. Several of analytical methods for the analysis of BPA have been reported including electrochemical processes, chemical oxidation, ozonization, spectrophotometric, chromatographic techniques. Compared with other conventional analytical techniques, optic sensors are reliable, providing quick results, low cost, easy to use, stands out as a much more advantageous method because of the high precision and sensitivity. In this work, a new photocured polymeric fluorescence sensor was prepared and characterized for Bisphenol-A (BPA) analysis. Characterization of the membrane was carried out by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) and Scanning Electron Microscope (SEM) techniques. The response characteristics of the sensor including dynamic range, pH effect and response time were systematically investigated. Acknowledgment: This work was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) under Grant 115Y469.

Keywords: Fluorescence, Photopolymerization, polymeric sensor, bisphenol-a

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16 Fluorescence-Based Biosensor for Dopamine Detection Using Quantum Dots

Authors: Sylwia Krawiec, Joanna Cabaj, Karol Malecha


Nowadays, progress in the field of the analytical methods is of great interest for reliable biological research and medical diagnostics. Classical techniques of chemical analysis, despite many advantages, do not permit to obtain immediate results or automatization of measurements. Chemical sensors have displaced the conventional analytical methods - sensors combine precision, sensitivity, fast response and the possibility of continuous-monitoring. Biosensor is a chemical sensor, which except of conventer also possess a biologically active material, which is the basis for the detection of specific chemicals in the sample. Each biosensor device mainly consists of two elements: a sensitive element, where is recognition of receptor-analyte, and a transducer element which receives the signal and converts it into a measurable signal. Through these two elements biosensors can be divided in two categories: due to the recognition element (e.g immunosensor) and due to the transducer (e.g optical sensor). Working of optical sensor is based on measurements of quantitative changes of parameters characterizing light radiation. The most often analyzed parameters include: amplitude (intensity), frequency or polarization. Changes in the optical properties one of the compound which reacts with biological material coated on the sensor is analyzed by a direct method, in an indirect method indicators are used, which changes the optical properties due to the transformation of the testing species. The most commonly used dyes in this method are: small molecules with an aromatic ring, like rhodamine, fluorescent proteins, for example green fluorescent protein (GFP), or nanoparticles such as quantum dots (QDs). Quantum dots have, in comparison with organic dyes, much better photoluminescent properties, better bioavailability and chemical inertness. These are semiconductor nanocrystals size of 2-10 nm. This very limited number of atoms and the ‘nano’-size gives QDs these highly fluorescent properties. Rapid and sensitive detection of dopamine is extremely important in modern medicine. Dopamine is very important neurotransmitter, which mainly occurs in the brain and central nervous system of mammals. Dopamine is responsible for the transmission information of moving through the nervous system and plays an important role in processes of learning or memory. Detection of dopamine is significant for diseases associated with the central nervous system such as Parkinson or schizophrenia. In developed optical biosensor for detection of dopamine, are used graphene quantum dots (GQDs). In such sensor dopamine molecules coats the GQD surface - in result occurs quenching of fluorescence due to Resonance Energy Transfer (FRET). Changes in fluorescence correspond to specific concentrations of the neurotransmitter in tested sample, so it is possible to accurately determine the concentration of dopamine in the sample.

Keywords: Quantum Dots, Biosensor, Fluorescence, Dopamine

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15 Characterization of Organic Matter in Spodosol Amazonian by Fluorescence Spectroscopy

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


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: Characterization, Humic Acid , Fluorescence, lifetime, Amazonian soil

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14 Highly Selective Polymeric Fluorescence Sensor for Cd(II) Ions

Authors: Soner Cubuk, M. Vezir Kahraman, Ece Kok Yetimoglu, Ozge Yilmaz


In this work, a polymer based highly selective fluorescence sensor membrane was prepared by the photopolymerization technique for the determination Cd(II) ion. Sensor characteristics such as effects of pH, response time and foreign ions on the fluorescence intensity of the sensor were also studied. Under optimized conditions, the polymeric sensor shows a rapid, stable and linear response for 4.45x10-⁹ mol L-¹ - 4.45x10-⁸ mol L-¹ Cd(II) ion with the detection limit of 6.23x10-¹⁰ mol L-¹. In addition, sensor membrane was selective which is not affected by common foreign metal ions. The concentrations of the foreign ions such as Pb²+, Co²+, Ag+, Zn²+, Cu²+, Cr³+ are 1000-fold higher than Cd(II) ions. Moreover, the developed polymeric sensor was successfully applied to the determination of cadmium ions in food and water samples. This work was supported by Marmara University, Commission of Scientific Research Project.

Keywords: Fluorescence, Photopolymerization, polymeric sensor, cadmium(II)

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13 Fractal Analysis of Polyacrylamide-Graphene Oxide Composite Gels

Authors: Önder Pekcan, Gülşen Akın Evingür


The fractal analysis is a bridge between the microstructure and macroscopic properties of gels. Fractal structure is usually provided to define the complexity of crosslinked molecules. The complexity in gel systems is described by the fractal dimension (Df). In this study, polyacrylamide- graphene oxide (GO) composite gels were prepared by free radical crosslinking copolymerization. The fractal analysis of polyacrylamide- graphene oxide (GO) composite gels were analyzed in various GO contents during gelation and were investigated by using Fluorescence Technique. The analysis was applied to estimate Df s of the composite gels. Fractal dimension of the polymer composite gels were estimated based on the power law exponent values using scaling models. In addition, here we aimed to present the geometrical distribution of GO during gelation. And we observed that as gelation proceeded GO plates first organized themselves into 3D percolation cluster with Df=2.52, then goes to diffusion limited clusters with Df =1.4 and then lines up to Von Koch curve with random interval with Df=1.14. Here, our goal is to try to interpret the low conductivity and/or broad forbidden gap of GO doped PAAm gels, by the distribution of GO in the final form of the produced gel.

Keywords: Scaling, Fluorescence, fractal, composite gels

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12 Fluorescence Quenching as an Efficient Tool for Sensing Application: Study on the Fluorescence Quenching of Naphthalimide Dye by Graphene Oxide

Authors: Shohre Rouhani, Sanaz Seraj


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, quenching, Graphene Oxide, naphthalimide dye

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11 Comparison of the Indocyanine Green Dye Method versus the Combined Method of Indigo Carmine Blue Dye with Indocyanine Green Fluorescence Imaging for Sentinel Lymph Node Biopsy in Breast Conservative Therapy for Early Breast Cancer

Authors: Nobuyuki Takemoto, Ai Koyanagi, Masanori Yasuda, Hiroshi Yamamoto


Background: Fluorescence imaging (FI) is one of the methods to identify sentinel lymph nodes (SLNs). However, the procedure is technically complicated and requires procedural skills, as SLN biopsy must be conducted in dim light conditions. As an improved version of this method, we introduced a combined method (Combined mixed dye and fluorescence; CMF) consisting of indigo carmine blue dye and FI. The direct visualization of SLNs under shadowless surgical light conditions is facilitated by the addition of the blue dye. We compared the SLN detection rates of CMF with that of the indocyanine green (ICG) dye method (ICG-D). Methods: A total of 202 patients with stage ≤ IIA breast cancer who underwent breast conservative therapy with separate incision from January 2004 to February 2017 were reviewed. Details of the two methods are as follows: (1) ICG-D: 2ml of ICG (10mg) was used and the green-stained SLNs were resected via a 3-4cm axillary incision; (2) CMF: A combination of 1ml of ICG (5mg) and 1-3ml of indigo carmine (4-12mg) was used. Using Photodynamic Eye (PDE), a 1.5-2 cm incision was made near the point of disappearance of the fluorescence and SLNs with intermediate color of blue and green were resected. Results: There were 92 ICG-D and 110 CMF cases. CMF resulted in a significantly higher detection rate than ICG-D (96.4% vs. 83.7%; p=0.003). This difference was particularly notable in those aged ≥ 60 years (98.3% vs. 74.3%) and individuals with BMI ≥ 25kg/m2 (90.3% vs. 58.3%). Conclusion: CMF is an effective method to identify SLNs which is safe, efficient, and cost-effective. Furthermore, radiation exposure can be avoided, and it can be performed in institutes without nuclear medicine facilities. CMF achieves a high SLN identification rate, and most of this procedure is feasible under shadowless surgical light conditions. CMF can reliably perform SLN biopsy even in those aged ≥ 60 years and individuals with BMI ≥ 25 kg/m2.

Keywords: Fluorescence, indigocarmine, sentinel lymph node biopsy, identification rate, indocyanine green (ICG)

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10 Comparative Analysis of Short and Long Term Salt Stress on the Photosynthetic Apparatus and Chloroplast Ultrastructure of Thellungiella salsuginea

Authors: Chedly Abdelly, Rahma Goussi, Walid Derbali, Arafet Manaa, Simone Cantamessa, Graziella Berta, Roberto Barbato


Salinity is one of the most important abiotic affecting plant growth and productivity worldwide. Photosynthesis, together with cell growth, is among the primary processes to be affected by salinity. Here, we report the effects of salinity stress on the primary processes of photosynthesis in a model halophyte Thellungiella Salsuginea. Plants were cultivated in hydroponic system with different NaCl concentrations (0, 100, 200 and 400 mM) during 2 weeks. The obtained results showed an obvious change in the photosynthetic efficiency of photosystem I (PSI) and phostosytem II (PSII), related to NaCl concentration supplemented to the medium and the stress duration considered. With moderate salinity (100 and 200 mM NaCl), no significant variation was observed in photosynthetic parameters of PSI and PSII and Chl fluorescence whatever the time of stress application. Also, the photosynthesis apparatus Fo, Fm and Fv fluorescence, as well as Fv/Fm were not affected by salt stress. While a significant decrease was observed on quantum yields Y(I), Y(II) and electron transport rate ETR(I), ETR(II) under high salt treatment (400 mM NaCl) with prolonged period (15 days). This reduction is quantitatively compensated by a corresponding increase of energy dissipation Y(NPQ) and a progressive decrease in Fv/Fm under salt treatment. The intensity of the OJIP fluorescence transient decreased with increase in NaCl concentration, with a major effect observed during prolonged period of salt stress. Ultrastructural analysis with Light Microscopy and Transmission Electron Microscopy of T. salsuginea chloroplasts showed some cellular changes, such as the shape of the mesophyll cells and number of chloroplast/cell only under higher NaCl concentration. Salt-stress caused the swelling of thylakoids in T. Salsuginea mesophyll with more accumulation of starch as compared to control plant.

Keywords: Photosynthesis, Fluorescence, Salt Stress, halophyte

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9 Fluorescent Analysis of Gold Nanoclusters-Wool Keratin Addition to Copper Ions

Authors: Yao Xing, Hong Ling Liu, Wei Dong Yu


With the increase of global population, it is of importance for the safe water supply, while, the water-monitoring method with the capability of rapidness, low-cost, green and robustness remains unsolved. In this paper, gold nanoclusters-wool keratin is added into copper ions measured by fluorescent method in order to probe copper ions in aqueous solution. The fluorescent results show that gold nanoclusters-wool keratin exhibits high stability of pHs, while it is sensitive to temperature and time. Based on Gauss fitting method, the results exhibit that the slope of gold nanoclusters-wool keratin with pH resolution under acidic condition is higher compared to it under alkaline solutions. Besides, gold nanoclusters-wool keratin added into copper ions shows a fluorescence turn-off response transferring from red to blue under UV light, leading to the dramatically decreased fluorescent intensity of gold nanoclusters-wool keratin solution located at 690 nm. Moreover, the limited concentration of copper ions tested by gold nanoclusters-wool keratin system is around 1 µmol/L, which meets the need of detection standards. The fitting slope of Stern-Volmer plot at low concentration of copper ions is larger than it at high concentrations, which indicates that aggregated gold nanoclusters are from small amounts to large numbers with the increasing concentration of copper ions. It is expected to provide novel method and materials for copper ions testing with low cost, high efficiency, and easy operability.

Keywords: Fluorescence, copper ions, gold nanoclusters, wool keratin

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8 Nano-Particle of π-Conjugated Polymer for Near-Infrared Bio-Imaging

Authors: Hiroyuki Aoki


Molecular imaging has attracted much attention recently, which visualizes biological molecules, cells, tissue, and so on. Among various in vivo imaging techniques, the fluorescence imaging method has been widely employed as a useful modality for small animals in pre-clinical researches. However, the higher signal intensity is needed for highly sensitive in vivo imaging. The objective of the current study is the development of a fluorescent imaging agent with high brightness for the tumor imaging of a mouse. The strategy to enhance the fluorescence signal of a bio-imaging agent is the increase of the absorption of the excitation light and the fluorescence conversion efficiency. We developed a nano-particle fluorescence imaging agent consisting of a π-conjugated polymer emitting a fluorescence signal in a near infrared region. A large absorption coefficient and high emission intensity at a near infrared optical window for biological tissue enabled highly sensitive in vivo imaging with a tumor-targeting ability by an EPR (enhanced permeation and retention) effect. The signal intensity from the π-conjugated fluorescence imaging agent is larger by two orders of magnitude compared to a quantum dot, which has been known as the brightest imaging agent. The π-conjugated polymer nano-particle would be a promising candidate in the in vivo imaging of small animals.

Keywords: Fluorescence, conjugated polymer, in vivo imaging, nano-particle, near-infrared

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7 A Study on Real-Time Fluorescence-Photoacoustic Imaging System for Mouse Thrombosis Monitoring

Authors: Chul Gyu Song, Moung Young Lee, Sang Hun Park, Su Min Yu, Hyun Sang Jo, Ji Hyeon Kim


A near-infrared light source used as a light source in the fluorescence imaging system is suitable for use in real-time during the operation since it has no interference in surgical vision. However, fluorescence images do not have depth information. In this paper, we configured the device with the research on molecular imaging systems for monitoring thrombus imaging using fluorescence and photoacoustic. Fluorescence imaging was performed using a phantom experiment in order to search the exact location, and the Photoacoustic image was in order to detect the depth. Fluorescence image obtained when evaluated through current phantom experiments when the concentration of the contrast agent is 25μg / ml, it was confirmed that it looked sharper. The phantom experiment is has shown the possibility with the fluorescence image and photoacoustic image using an indocyanine green contrast agent. For early diagnosis of cardiovascular diseases, more active research with the fusion of different molecular imaging devices is required.

Keywords: Fluorescence, carotid artery, photoacoustic, indocyanine green

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6 Spectrofluorimetric Investigation of Copper (II), Cobalt (II), Calcium (II), and Ferric (III) Influence on the Ciprofloxacin Binding to Bovine Serum Albumin

Authors: Ahmed K. Youssef, Shawkat M. B. Aly


The interaction between ciprofloxacin and bovine serum albumin (BSA) was investigated by UV-Visible absorption and fluorescence spectroscopy. The influence of Cu²⁺ Ca²⁺, Co²⁺, and Fe³⁺ on the Cip-BSA interaction was investigated. The quenching of the BSA fluorescence emission in presence of ciprofloxacin as well as the influence of metal ions on the interaction was analyzed using the Stern-Volmer equation. The Stern-Volmer quenching constant, Kₛᵥ was calculated in presence and absence of the metal ions at the physiological pH of 7.4 using phosphate buffer. The experimental results showed that interaction mainly static in nature and quenching rate constant is decreased in presence of the studied metal ions with exception of Cu²⁺ ions. The decrease observed in the Kₛᵥ values in presence of Co²⁺, Ca²⁺, and Fe³⁺ can be understood on basis of competition between these metal and Cip when both of them existed in the BSA solution. Cu²⁺ induces interaction between Cip and BSA at faster quenching rates as inferred from the observed increase in the Kₛᵥ value. This allowed us to propose that copper (II) ions are directly involved in the process of Cip binding to BSA. The binding constant for Cip on BSA was determined and the metal ions effect on it was examined as well and their values were in line with the Kₛᵥ values.

Keywords: Fluorescence, ciprofloxacin, bovine serum albumin, metal ions effect

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5 Ionic Liquid and Chemical Denaturants Effects on the Fluorescence Properties of the Laccase

Authors: Othman Saoudi


In this work, we have interested in the investigation of the chemical denaturants and synthesized ionic liquids effects on the fluorescence properties of the laccase from Trametes versicolor. The fluorescence properties of the laccase result from the presence of Tryptophan, which has an aromatic core responsible for the absorption in ultra violet domain and the emission of the photons of fluorescence. The effect Pyrrolidinuim Formate ([pyrr][F]) and Morpholinium Formate ([morph][F]) ionic liquids on the laccase behavior for various volumetric fractions are studied. We have shown that the fluorescence spectrum relative to the [pyrr][F] presents a single band with a maximum around 340 nm and a secondary peak at 361 nm for a volumetric fraction of 20% v/v. For concentration superiors to 40%, the fluorescence intensity decreases and a displacement of the peaks toward higher wavelengths has occurred. For the [morph][F], the fluorescence spectrum showed a single band around 340 nm. The intensity of the principal peak decreases for concentration superiors to 20% v/v. From the plot representing the variation of the λₘₐₓ versus the volumetric concentration, we have determined the concentration of the half-transitions C1/2. These concentrations are equal to 42.62% and 40.91% v/v in the presence of [pyrr][F] and [morph][F] respectively. For the chemical denaturation, we have shown that the fluorescence intensity decreases with increasing denaturant concentrations where the maximum of the wavelength of emission shifts toward the higher wavelengths. We have also determined from the spectrum relative to the urea and GdmCl, the unfolding energy, ∆GD. The results show that the variation of the unfolding energy as a function of the denaturant concentrations varies according to the linear regression model. We have demonstrated also that the half-transitions C1/2 have occurred for urea and GdmCl denaturants concentrations around 3.06 and 3.17 M respectively.

Keywords: Ionic Liquids, Fluorescence, laccase, chemical denaturants

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4 Peptide-Gold Nanocluster as an Optical Biosensor for Glycoconjugate Secreted from Leishmania

Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino


In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and [email protected], therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the [email protected] conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the [email protected] To our best knowledge the fabrication of an optical biosensor based on [email protected] for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: Biosensing, Fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

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3 A Dual Channel Optical Sensor for Norepinephrine via Situ Generated Silver Nanoparticles

Authors: Shalini Menon, K. Girish Kumar


Norepinephrine (NE) is one of the naturally occurring catecholamines which act both as a neurotransmitter and a hormone. Catecholamine levels are used for the diagnosis and regulation of phaeochromocytoma, a neuroendocrine tumor of the adrenal medulla. The development of simple, rapid and cost-effective sensors for NE still remains a great challenge. Herein, a dual-channel sensor has been developed for the determination of NE. A mixture of AgNO₃, NaOH, NH₃.H₂O and cetrimonium bromide in appropriate concentrations was taken as the working solution. To the thoroughly vortexed mixture, an appropriate volume of NE solution was added. After a particular time, the fluorescence and absorbance were measured. Fluorescence measurements were made by exciting at a wavelength of 400 nm. A dual-channel optical sensor has been developed for the colorimetric as well as the fluorimetric determination of NE. Metal enhanced fluorescence property of nanoparticles forms the basis of the fluorimetric detection of this assay, whereas the appearance of brown color in the presence of NE leads to colorimetric detection. Wide linear ranges and sub-micromolar detection limits were obtained using both the techniques. Moreover, the colorimetric approach was applied for the determination of NE in synthetic blood serum and the results obtained were compared with the classic high-performance liquid chromatography (HPLC) method. Recoveries between 97% and 104% were obtained using the proposed method. Based on five replicate measurements, relative standard deviation (RSD) for NE determination in the examined synthetic blood serum was found to be 2.3%. This indicates the reliability of the proposed sensor for real sample analysis.

Keywords: Colorimetry, Fluorescence, Silver Nanoparticles, norepinephrine

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2 Unveiling the Self-Assembly Behavior and Salt-Induced Morphological Transition of Double PEG-Tailed Unconventional Amphiphiles

Authors: Rita Ghosh, Joykrishna Dey


PEG-based amphiphiles are of tremendous importance for its widespread applications in pharmaceutics, household purposes, and drug delivery. Previously, a number of single PEG-tailed amphiphiles having significant applications have been reported from our group. Therefore, it was of immense interest to explore the properties and application potential of PEG-based double tailed amphiphiles. Herein, for the first time, two novel double PEG-tailed amphiphiles having different PEG chain lengths have been developed. The self-assembly behavior of the newly developed amphiphiles in aqueous buffer (pH 7.0) was thoroughly investigated at 25 oC by a number of techniques including, 1H-NMR, and steady-state and time-dependent fluorescence spectroscopy, dynamic light scattering, transmission electron microscopy, atomic force microscopy, and isothermal titration calorimetry. Despite having two polar PEG chains both molecules were found to have strong tendency to self-assemble in aqueous buffered solution above a very low concentration. Surprisingly, the amphiphiles were shown to form stable vesicles spontaneously at room temperature without any external stimuli. The results of calorimetric measurements showed that the vesicle formation is driven by the hydrophobic effect (positive entropy change) of the system, which is associated with the helix-to-random coil transition of the PEG chain. The spectroscopic data confirmed that the bilayer membrane of the vesicles is constituted by the PEG chains of the amphiphilic molecule. Interestingly, the vesicles were also found to exhibit structural transitions upon addition of salts in solution. These properties of the vesicles enable them as potential candidate for drug delivery.

Keywords: Microscopy, Fluorescence, PEG, vesicles, Double-tailed amphiphiles

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1 Synthesis of Fluorescent PET-Type “Turn-Off” Triazolyl Coumarin Based Chemosensors for the Sensitive and Selective Sensing of Fe⁺³ Ions in Aqueous Solutions

Authors: Aidan Battison, Neliswa Mama


Environmental pollution by ionic species has been identified as one of the biggest challenges to the sustainable development of communities. The widespread use of organic and inorganic chemical products and the release of toxic chemical species from industrial waste have resulted in a need for advanced monitoring technologies for environment protection, remediation and restoration. Some of the disadvantages of conventional sensing methods include expensive instrumentation, well-controlled experimental conditions, time-consuming procedures and sometimes complicated sample preparation. On the contrary, the development of fluorescent chemosensors for biological and environmental detection of metal ions has attracted a great deal of attention due to their simplicity, high selectivity, eidetic recognition, rapid response and real-life monitoring. Coumarin derivatives S1 and S2 (Scheme 1) containing 1,2,3-triazole moieties at position -3- have been designed and synthesized from azide and alkyne derivatives by CuAAC “click” reactions for the detection of metal ions. These compounds displayed a strong preference for Fe3+ ions with complexation resulting in fluorescent quenching through photo-induced electron transfer (PET) by the “sphere of action” static quenching model. The tested metal ions included Cd2+, Pb2+, Ag+, Na+, Ca2+, Cr3+, Fe3+, Al3+, Cd2+, Ba2+, Cu2+, Co2+, Hg2+, Zn2+ and Ni2+. The detection limits of S1 and S2 were determined to be 4.1 and 5.1 uM, respectively. Compound S1 displayed the greatest selectivity towards Fe3+ in the presence of competing for metal cations. S1 could also be used for the detection of Fe3+ in a mixture of CH3CN/H¬2¬O. Binding stoichiometry between S1 and Fe3+ was determined by using both Jobs-plot and Benesi-Hildebrand analysis. The binding was shown to occur in a 1:1 ratio between the sensor and a metal cation. Reversibility studies between S1 and Fe3+ were conducted by using EDTA. The binding site of Fe3+ to S1 was determined by using 13 C NMR and Molecular Modelling studies. Complexation was suggested to occur between the lone-pair of electrons from the coumarin-carbonyl and the triazole-carbon double bond.

Keywords: Fluorescence, triazole, coumarin, chemosensor, click" chemistry, static quenching

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