Evaluation of fluorescent dye degradation indirectly induced by x-ray ionizing radiation.

This work evaluated the fluorescent dye degradation indirectly induced by ionizing radiation with high energy photons (50 keV). Aqueous gels of agarose with low concentrations of Rhodamine 6G and Fluorescein were submitted to doses of x-ray radiation up to 200 Gy. The dye degradation was analyzed by fluorescence spectroscopy, using an excitation light-emitting diode with a peak wavelength of 462 nm. A rate equation model of fluorophores and radicals' species populations was developed to describe the degradation time behavior of the fluorescent solutions. The model suggests fluorescent dyes should be used in dosimetry.

Effectiveness of varnish with CPP-ACP in prevention of caries lesions around orthodontic brackets: an OCT evaluation.

OBJECTIVE: To evaluate the in vitro efficiency of applying varnish containing casein phosphopeptide (CPP) and amorphous calcium phosphate (ACP) in prevention of caries lesions around orthodontic brackets. MATERIALS AND METHODS: For this purpose, brackets were bonded to the vestibular surface of bovine incisors, and eight groups were formed (n = 15) according to exposure of oral hygiene substances and enamel varnish: 1 (control) brushing only performed, 2 (control) brushing + use of mouth wash with fluoride, 3 Duraphat varnish application only (Colgate-Palmolive Ind. E Com. Ltda, São Paulo, SP, Brazil), 4 Duraphat + brushing, 5 Duraphat + brushing + mouth wash, 6 MI Varnish application (GC America, USA), 7 MI + brushing, and 8 MI + brushing + mouth wash. The experimental groups alternated between pH cycling and the procedures described and were kept in an oven at temperature of 37°C. Both brushing and immersion in solutions was performed in a time interval of 1 minute, followed by washing in deionized water three times a day for 28 days of experimentation. Afterwards, evaluation by optical coherence tomography (OCT) of the special type (Ganymede OCT/Thorlabs, Newton, USA) was performed. In each group, a scanning exam of the white spot lesion area (around the region where brackets were bonded) and depth measurement of caries lesions were performed. RESULTS: Groups 1 and 3 were shown to differ statistically from groups 5, 6, 7, and 8 (p = 0.000). MI Varnish was shown to be more effective in diminishing caries lesion depth, compared with Duraphat, irrespective of being associated with brushing and mouth wash, or not. LIMITATIONS: The major limitation of this study is that it is a study in which demineralization was obtained with the use of chemical products, and did not occur due to the presence of Streptococcus mutans and its acid byproducts. CONCLUSION: Application CPP-ACP-containing varnish irrespective of being associated with brushing and mouthwash, or not, reduced depth of caries lesions around orthodontic brackets.

Dengue immunoassay with an LSPR fiber optic sensor.

Dengue fever is a viral disease that affects millions of people worldwide. Specific tests for dengue are not usually performed due to high costs, complicated procedures and, in some cases, long time to yield a result. For widespread use of specific tests to be possible, fast, reliable and fairly simple methods are needed. In this paper, we present a new dengue diagnostic method for the acute phase of the infection. The method proposed uses an all-optical fiber sensor based on Localized Surface Plasmon Resonance (LSPR) and specular reflection from gold nanoparticles (AuNPs). Dengue anti-NS1 antibody was immobilized on AuNPs deposited on the endface of a standard multimode fiber (62.5 µm/125 µm). The sensor is able to detect NS1 antigen at different concentrations, with limit of quantification estimated to be 0.074 µg/ml = 1.54 nM. These results indicate that the sensor could potentially be used for dengue diagnosis in the acute phase of the infection.

Singlet oxygen generation enhanced by silver-pectin nanoparticles.

We demonstrate the potential application of silver-pectin nanoparticles on photodynamic therapy, on a solution-base platform. Photodynamic therapy is a medical technique which uses a combination of photosensitizing drugs and light to induce selective damage on the target tissue, by electronically excited and highly reactive singlet state of oxygen. Metal enhanced singlet oxygen generation in riboflavin water solution with silver-pectin nanoparticles was observed and quantified. Here 13 nm silver nanospheres enclosed by a pectin layer were synthesized and it interaction with riboflavin molecule was analyzed. Pectin, a complex carbohydrate found in plants primary cell walls, was used to increase the biocompatibility of the silver nanoparticles and to improve metal enhanced singlet oxygen generation (28.5 %) and metal-enhanced fluorescence (30.7 %) processes at room temperature. The singlet oxygen sensor fluorescent green reagent was used to quantify the enhancement of the riboflavin singlet oxygen production induced by the silver colloid. We report a 1.7-fold increase of riboflavin emission and a 1.8-fold enhancement of singlet oxygen production.

Selecting High-Performance Gold Nanorods for Photothermal Conversion.

In this work, we establish a new paradigm on identifying optimal arbitrarily shaped metallic nanostructures for photothermal applications. Crucial thermo-optical parameters that rule plasmonic heating are appraised, exploring a nanoparticle size-dependence approach. Our results indicate two distinct figures of merit for the optimization of metallic nanoheaters, under both non-cumulative femtosecond and continuum laser excitation. As a case study, gold nanorods are evaluated for infrared photothermal conversion in water, and the influence of the particle length and diameter are depicted. For non-cumulative femtosecond pulses, efficient photothermal conversion is observed for gold nanorods of small volumes. For continuous wave (CW) excitation at 800 nm and 1064 nm, the optimal gold nanorod dimensions (in water) are, respectively, 90 × 25nm and 150 × 30 nm. Figure of Merit (FoM) variations up to 700% were found considering structures with the same peak wavelength. The effect of collective heating is also appraised. The designing of high-performance plasmonic nanoparticles, based on quantifying FoM, allows a rational use of nanoheaters for localized photothermal applications.

Optimizing and Quantifying Gold Nanospheres Based on LSPR Label-Free Biosensor for Dengue Diagnosis.

The localized surface plasmon resonance (LSPR) due to light-particle interaction and its dependence on the surrounding medium have been widely manipulated for sensing applications. The sensing efficiency is governed by the refractive index-based sensitivity (ηRIS) and the full width half maximum (FWHM) of the LSPR spectra. Thereby, a sensor with high precision must possess both requisites: an effective ηRIS and a narrow FWHM of plasmon spectrum. Moreover, complex nanostructures are used for molecular sensing applications due to their good ηRIS values but without considering the wide-band nature of the LSPR spectrum, which decreases the detection limit of the plasmonic sensor. In this article, a novel, facile and label-free solution-based LSPR immunosensor was elaborated based upon LSPR features such as extinction spectrum and localized field enhancement. We used a 3D full-wave field analysis to evaluate the optical properties and to optimize the appropriate size of spherical-shaped gold nanoparticles (Au NPs). We found a change in Au NPs' radius from 5 nm to 50 nm, and an increase in spectral resonance peak depicted as a red-shift from 520 nm to 552 nm. Using this fact, important parameters that can be attributed to the LSPR sensor performance, namely the molecular sensitivity, FWHM, ηRIS, and figure of merit (FoM), were evaluated. Moreover, computational simulations were used to assess the optimized size (radius = 30 nm) of Au NPs with high FoM (2.3) and sharp FWHM (44 nm). On the evaluation of the platform as a label-free molecular sensor, Campbell's model was performed, indicating an effective peak shift in the adsorption of the dielectric layer around the Au NP surface. For practical realization, we present an LSPR sensor platform for the identification of dengue NS1 antigens. The results present the system's ability to identify dengue NS1 antigen concentrations with the limit of quantification measured to be 0.07 µg/mL (1.50 nM), evidence that the optimization approach used for the solution-based LSPR sensor provides a new paradigm for engineering immunosensor platforms.

Identifying high performance gold nanoshells for singlet oxygen generation enhancement.

Metallic nanostructures can improve the production of singlet oxygen (1O2) of a photosensitizer during photodynamic therapy (PDT) . Engineering a high performance nanoparticle is mandatory for an appropriate use of plasmonic nanostructures in PDT. Metal enhanced singlet oxygen generation requires the use of nanoparticles with high scattering efficiency, capable of inducing a significant electric field enhancement and with plasmon peak overlapping the photosensitizer absorption spectrum. Herein, we report the optimization of nanoshells structure (silica core radius and gold shell thickness) to increase the singlet oxygen production by Methylene Blue photosensitizer. A 3D Full-wave field analysis was used to evaluate the plasmonic spectrum, scattering efficiency and localized field intensity of Au nanoshells as a function of their dimensions. The 40/20 core radius/shell thickness optimized gold nanoshell showed 75% scattering efficiency and field enhancement up to 35 times. Metal-enhanced singlet oxygen generation was observed and quantified for Methylene Blue water solution with gold nanoshell particles. Moreover, the influence of the irradiation time and the metallic nanostructures concentration on metal enhanced singlet oxygen generation were also appraised. The experimental results showed that the use of gold nanoshell improved 320% the 1O2 production in a MB solution. The approach used to select a high performance metallic nanoparticle provides insights on engineering plasmonic structures for metal enhanced singlet oxygen generation for PDT.

Absolute and Relative Methods for Fluorescence Quantum Yield Evaluation of Quantum Dots.

Optical spectroscopy techniques are crucial for the evaluation and use of quantum dots (QDs) in life and materials science. In that context, the fluorescence quantum yield (Φf) is an essential parameter in the assessment of the luminescent features of QDs. The fluorescence quantum yield can be defined as the ratio of the number of emitted photons to the number of absorbed photons by a luminescent material. In this chapter, we describe absolute and relative methods to measure the fluorescence quantum yield of QDs in solution phase. The advantages and limitations of the techniques are reviewed.

Evaluating viscoelastic properties and membrane electrical charges of red blood cells with optical tweezers and cationic quantum dots - applications to ß-thalassemia intermedia hemoglobinopathy.

Biomechanical and electrical properties are important to the performance and survival of red blood cells (RBCs) in the microcirculation. This study proposed and explored methodologies based on optical tweezers and cationic quantum dots (QDs) as biophotonic tools to characterize, in a complementary way, viscoelastic properties and membrane electrical charges of RBCs. The methodologies were applied to normal (HbA) and ß-thalassemia intermedia (Hbß) RBCs. The ß-thalassemia intermedia disease is a hereditary hemoglobinopathy characterized by a reduction (or absence) of ß-globin chains, which leads to α-globin chains precipitation. The apparent elasticity (µ) and membrane viscosity (ηm) of RBCs captured by optical tweezers were obtained in just a single experiment. Besides, the membrane electrical charges were evaluated by flow cytometry, exploring electrostatic interactions between cationic QDs, stabilized with cysteamine, with the negatively charged RBC surfaces. Results showed that Hbß RBCs are less elastic, have a higher ηm, and presented a reduction in membrane electrical charges, when compared to HbA RBCs. Moreover, the methodologies based on optical tweezers and QDs, here proposed, showed to be capable of providing a deeper and integrated comprehension on RBC rheological and electrical changes, resulting from diverse biological conditions, such as the ß-thalassemia intermedia hemoglobinopathy.

TiO2 -coated fluoride nanoparticles for dental multimodal optical imaging.

Core-shell nanostructures associated with photonics techniques have found innumerous applications in diagnostics and therapy. In this work, we introduce a novel core-shell nanostructure design that serves as a multimodal optical imaging contrast agent for dental adhesion evaluation. This nanostructure consists of a rare-earth-doped (NaYF4 :Yb 60%, Tm 0.5%)/NaYF4 particle as the core (hexagonal prism, ~51 nm base side length) and the highly refractive TiO2 material as the shell (~thickness of 15 nm). We show that the TiO2 shell provides enhanced contrast for optical coherence tomography (OCT), while the rare-earth-doped core upconverts excitation light from 975 nm to an emission peaked at 800 nm for photoluminescence imaging. The OCT and the photoluminescence wide-field images of human tooth were demonstrated with this nanoparticle core-shell contrast agent. In addition, the described core-shell nanoparticles (CSNps) were dispersed in the primer of a commercially available dental bonding system, allowing clear identification of dental adhesive layers with OCT. We evaluated that the presence of the CSNp in the adhesive induced an enhancement of 67% scattering coefficient to significantly increase the OCT contrast. Moreover, our results highlight that the upconversion photoluminescence in the near-infrared spectrum region is suitable for image of deep dental tissue.

Photodynamic inactivation assisted by localized surface plasmon resonance of silver nanoparticles: In vitro evaluation on Escherichia coli and Streptococcus mutans.

Localized surface plasmon resonance (LSPR) of gold nanoparticles has been reported to increase the antimicrobial effect of the photodynamic therapy. Although silver nanoparticles (AgNPs) are an efficient growth inhibitor of microorganisms, no studies exploring LSPR of AgNPs to enhance the photodynamic inactivation (PDI) have been related. In this work, we described the LSPR phenomenon of AgNP sand investigated its interaction with riboflavin, a natural photosensitizer. We evaluated the use of AgNPs coated with pectin (p-AgNP) in riboflavin (Rb)-mediated PDI of Escherichia coli (Gram- bacteria) and Streptococcus mutans (Gram + bacteria) using a blue light-emitting diode (λ = 455 ±â€¯20 nm) of optical power 200 mW. Irradiance was 90 mW/cm2 and radiant exposure varied according to the time exposure. Uptake of Rb and p-AgNP by the cells was evaluated by measuring the supernatant absorption spectra of the samples. We observed that LSPR of p-AgNPs was able to enhance the riboflavin photodynamic action on S. mutans but not on E. coli, probably due to the lower uptake of Rb by E. coli. Taken together, our results provide insights to explore the use of the LPRS promoted by silver nanostructures to optimize antimicrobial PDI protocols.

Damage induced in red blood cells by infrared optical trapping: an evaluation based on elasticity measurements.

We evaluated the damage caused to optically trapped red blood cells (RBCs) after 1 or 2 min of exposure to near-infrared (NIR) laser beams at 785 or 1064 nm. Damage was quantified by measuring cell elasticity using an automatic, real-time, homemade, optical tweezer system. The measurements, performed on a significant number (hundreds) of cells, revealed an overall deformability decrease up to ∼104% after 2 min of light exposure, under 10 mW optical trapping for the 785-nm wavelength. Wavelength dependence of the optical damage is attributed to the light absorption by hemoglobin. The results provided evidence that RBCs have their biomechanical properties affected by NIR radiation. Our findings establish limits for laser applications with RBCs.

Automatic real time evaluation of red blood cell elasticity by optical tweezers.

Optical tweezers have been used to trap, manipulate, and measure individual cell properties. In this work, we show that the association of a computer controlled optical tweezers system with image processing techniques allows rapid and reproducible evaluation of cell deformability. In particular, the deformability of red blood cells (RBCs) plays a key role in the transport of oxygen through the blood microcirculation. The automatic measurement processes consisted of three steps: acquisition, segmentation of images, and measurement of the elasticity of the cells. An optical tweezers system was setup on an upright microscope equipped with a CCD camera and a motorized XYZ stage, computer controlled by a Labview platform. On the optical tweezers setup, the deformation of the captured RBC was obtained by moving the motorized stage. The automatic real-time homemade system was evaluated by measuring RBCs elasticity from normal donors and patients with sickle cell anemia. Approximately 150 erythrocytes were examined, and the elasticity values obtained by using the developed system were compared to the values measured by two experts. With the automatic system, there was a significant time reduction (60×) of the erythrocytes elasticity evaluation. Automated system can help to expand the applications of optical tweezers in hematology and hemotherapy.

Quantum dots fluorescence quantum yield measured by Thermal Lens Spectroscopy.

An essential parameter to evaluate the light emission properties of fluorophores is the fluorescence quantum yield, which quantify the conversion efficiency of absorbed photons to emitted photons. We detail here an alternative nonfluorescent method to determine the absolute fluorescence quantum yield of quantum dots (QDs). The method is based in the so-called Thermal Lens Spectroscopy (TLS) technique, which consists on the evaluation of refractive index gradient thermally induced in the fluorescent material by the absorption of light. Aqueous dispersion carboxyl-coated cadmium telluride (CdTe) QDs samples were used to demonstrate the Thermal Lens Spectroscopy technical procedure.

In situ gold nanoparticles formation: contrast agent for dental optical coherence tomography.

In this work we demonstrate the potential use of gold nanoparticles as contrast agents for the optical coherence tomography (OCT) imaging technique in dentistry. Here, a new in situ photothermal reduction procedure was developed, producing spherical gold nanoparticles inside dentinal layers and tubules. Gold ions were dispersed in the primer of commercially available dental bonding systems. After the application and permeation in dentin by the modified adhesive systems, the dental bonding materials were photopolymerized concurrently with the formation of gold nanoparticles. The gold nanoparticles were visualized by scanning electron microscopy (SEM). The SEM images show the presence of gold nanospheres in the hybrid layer and dentinal tubules. The diameter of the gold nanoparticles was determined to be in the range of 40 to 120 nm. Optical coherence tomography images were obtained in two- and three-dimensions. The distribution of nanoparticles was analyzed and the extended depth of nanosphere production was determined. The results show that the OCT technique, using in situ formed gold nanoparticles as contrast enhancers, can be used to visualize dentin structures in a non-invasive and non-destructive way.

Nonlinear excitation of tryptophan emission enhanced by silver nanoparticles.

We measured and analyzed the behavior of the fluorescence of tryptophan water solutions with and without silver nanoparticles, excited by one, two and three photon processes. Two different colloids with silver nanoparticles with distinct diameters (0.65 nm and 9 nm) were used in the experiments. Fluorescence quenching was observed with one and two photon excitation. However, upon three-photon excitation, significant fluorescence enhancement was observed in the colloid. In this case excitation of the amino acid is assisted by the nonlinear absorption of infrared light by the silver nanoparticles. In this paper we are proposing a new way to explore metallic nanoparticles to enhance autofluorescence of biomolecules.