RESUMO
Peanuts are among the most common food allergies, which may result in life-threatening reactions in certain people. For this reason, it is very important to monitor the presence of peanuts in the food system. Biosensors are an emerging way of detecting allergen proteins. In this research, we present a surface enhanced Raman spectroscopy (SERS) technique to detect the main allergen protein, Ara h1. The sensors were biodegradable and made out of a corn protein, zein. Nanophotonic structures on zein films consisted of gold coated pyramid structures. It was found that both detection and quantification was possible by using a statistical clustering technique principal component analysis (PCA). An optimization in data processing yielded the result that baseline correction and shorter data collection times were needed in order to successfully cluster data. The limit of detection was found to be 0.14 mg/ml. Furthermore, specificity of the sensor was provided by functionalizing the surface with monoclonal antibodies of Ara h1. Antibody functionalization, and Ara h1 capturing was tested and identified by also utilizing PCA analysis. As a proof-of-concept, this study showed that a biodegradable platform can be used in detection of a peanut allergen protein, Ara h1, using surface enhanced Raman spectroscopy.
Assuntos
Antígenos de Plantas/análise , Técnicas Biossensoriais/métodos , Glicoproteínas/análise , Ouro/química , Nanotecnologia/métodos , Proteínas de Plantas/análise , Análise Espectral Raman/métodos , Zeína/química , Anticorpos Monoclonais/imunologia , Antígenos de Plantas/imunologia , Glicoproteínas/imunologia , Proteínas de Membrana , Proteínas de Plantas/imunologia , Propriedades de SuperfícieRESUMO
Zein has been widely studied as a biopolymer due to its unique film-forming abilities. Surface properties are of high importance for certain applications which include microfluidics and tissue engineering, as they drastically affect the end result. It is important to develop techniques to modify zein surface properties without compromising bulk material properties. In this study, we developed a facile technique to change the water affinity of zein film surfaces, compatible with patterning techniques via soft lithography. This is achieved by a simple solvent casting technique onto a polydimethylsilohexane (PDMS) substrate that was exposed to oxygen plasma. Water contact angle measurements (WCA) were used to assess the hydrophillicity of zein surfaces and they reached as low as 20°. Atomic force microscopy, optical absorbance and light microscopy were used to study the characteristics of the film and its surface topography. Hydrophilic zein surfaces had higher roughness values compared to hydrophobic ones. Surface roughness, introduced by sandpaper and gratings does not have the same effect as surface chemistry. The amphiphilic nature of plasticizer oleic acid also contributed to the change in the water contact angle of the films. In conclusion, we demonstrated that zein film's surface properties can be controlled by its ability to self-assemble depending on the substrate that it is being cast on.
Assuntos
Dimetilpolisiloxanos/química , Ácido Oleico/química , Oxigênio/química , Zeína/química , Interações Hidrofóbicas e Hidrofílicas , Microscopia de Força Atômica , Plastificantes/química , Propriedades de SuperfícieRESUMO
We have implemented a multifunctional optofluidic sensor that can monitor changes in the refractive index and pressure of biofluid simultaneously and can detect free-solution molecular interaction in situ. In this Letter, we demonstrate two major improvements of this sensor proven by both simulation and experiments. One improvement is the broader measurement range of refractive index by making the diffraction grating with high-index polymer. The other improvement is the separation of refractive index sensing from opacity sensing by using the relative power ratio of diffraction orders. This simple, compact and low-cost multifunctional optofluidic sensor has the potential to be used for in situ biofluid monitoring.
Assuntos
Técnicas Analíticas Microfluídicas/instrumentação , Fenômenos Ópticos , Técnicas Analíticas Microfluídicas/economia , Sacarose/análiseRESUMO
We present an optofluidic sensor based on an elastomeric two-dimensional (2D) grating integrated inside a hemispherical fluid chamber. A laser beam is diffracted before (reflection) and after (transmission) going through the grating and liquid in the dome chamber. The sensing mechanism is investigated and simulated with a finite-difference time-domain-based electromagnetic method. For the experiment, by analyzing the size, power, and shape of the 2D diffraction patterns, we can retrieve multiple parameters of the liquid, including the refractive index, pressure, and opacity with high sensitivity. We demonstrate that the glucose concentration can be monitored when mixed in a different concentrated phosphate-buffered saline solution. The free-solution binding of bovine serum albumin (BSA) and anti-BSA IgG is detected with this optical sensor. This low-cost, multifunctional, and reliable optofluidic sensor has the potential to be used as a monitor of biofluid, such as blood in hemodialysis.
Assuntos
Elastômeros/química , Microfluídica , Animais , Glicemia/análise , Bovinos , Simulação por Computador , Desenho de Equipamento , Glucose/análise , Imunoglobulina G/análise , Óptica e Fotônica , Fosfatos/química , Pressão , Refratometria , Diálise Renal , Sais/química , Soroalbumina Bovina/análiseRESUMO
Gelatin nanoparticles coated with Cathepsin D-specific peptides were developed as a vehicle for the targeted delivery of the cancer drug doxorubicin (DOX) to treat breast malignancy. Cathepsin D, a breast cancer cell secretion enzyme, triggered the release of DOX by digesting the protective peptide-coating layer of nanoparticles. Fabricated nanoparticles were successfully detected with ultrasound imaging in both in vitro conditions and in vivo mouse cancer models. Cell viability experiments were conducted to determine the efficacy of biomarker activation specific to breast cancer cell lines. These experimental results were compared with the outcome of a viability experiment conducted on noncancerous cells. Viability decreased in human MCF7 mammary adenocarcinoma and mouse 4T1 mammary carcinoma cells, while that of noncancerous 3T3 fibroblast cells remained unaffected. Next, a real-time video of nanoparticle flow in mouse models was obtained using in vivo ultrasound imaging. The fluorescent profile of DOX was used as a means to examine nanoparticle localization in vivo. Results show the distribution of nanoparticles concentrated primarily within bladder and tumor sites of subject mice bodies. These findings support the use of biomarker coated nanoparticles in target specific therapy for breast cancer treatment.
Assuntos
Antineoplásicos/farmacocinética , Biomarcadores Tumorais/metabolismo , Portadores de Fármacos/química , Nanopartículas/química , Animais , Antineoplásicos/química , Biomarcadores Tumorais/química , Neoplasias da Mama/química , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Doxorrubicina/química , Doxorrubicina/farmacocinética , Feminino , Humanos , Células MCF-7 , Camundongos , Neoplasias Experimentais/química , Neoplasias Experimentais/metabolismo , Polímeros/químicaRESUMO
Water disinfection byproducts (DBPs) are a class of chemicals that are produced when chemical disinfectants react with organic materials in untreated water. Cytotoxicity and genotoxicity of DBPs have been systematically evaluated to compile a comparative, quantitative database of in vitro mammalian cell toxicity of DBPs. However, one of the most challenging limitations for current DBP cytotoxicity assessment assays is sample availability. Although our current cytotoxicity assay using a 96-well microplate has been designed to reduce sample consumption, further minimization of the size of the test system would allow us to explore various possibilities for point-of-care applications. We have developed a microfluidic device with micro-pillars that shows high uniformity in distribution of cells across all chambers with low cell count. We compare the performance between the 96-well microplate and the microfluidic device by running 72-hour standalone-on-chip cell culture and cytotoxicity analysis experiments, using dimethyl sulfoxide (DMSO) and ethanol as model toxic agents, and bromoacetic acid (BAA) as a representative DBP. The results show close agreement between the two systems. The measured LC(50) values for the 96-well microplate and the microfluidic device are 1.54% v/v and 1.27% v/v for DMSO, 1.44% v/v and 2.92% v/v for ethanol, and 17.6 µM and 8.20 µM for BAA, respectively. The micro-pillar microfluidic device offers a great reduction in sample consumption while maintaining the accuracy of the cytotoxicity analyses of water disinfection byproducts.
Assuntos
Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Desinfecção , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Água/análise , Animais , Células CHO , Cricetinae , Cricetulus , Fatores de Tempo , Água/químicaRESUMO
We have fabricated nanotextured Si substrates that exhibit controllable optical reflection intensities and colors. Si nanopore has a photon trapping nanostructure but has abrupt changes in the index of refraction displaying a darkened specular reflection. Nanoscrew Si shows graded refractive-index photon trapping structures that enable diffuse reflection to be as low as 2.2% over the visible wavelengths. By tuning the 3D nanoscale silicon structure, the optical reflection peak wavelength and intensity are changed in the wavelength range of 300-800 nm, making the surface have different reflectivity and apparent colors. The relation between the surface optical properties with the spatial features of the photon trapping nanostructures is examined. Integration of photon trapping structures with planar Si structure on the same substrate is also demonstrated. The tunable photon trapping silicon structures have potential applications in enhancing the performance of semiconductor photoelectric devices.
RESUMO
One-dimensional nanostructures, such as nanowhisker, nanorod, nanowire, nanopillar, nanocone, nanotip, nanoneedle, have attracted significant attentions in the past decades owing to their numerous applications in electronics, photonics, energy conversion and storage, and interfacing with biomolecules and living cells. The manufacturing of nanostructured devices relies on either bottom-up approaches such as synthesis or growth process or top-down approaches such as lithography or etching process. Here we report a unique, synchronized, and simultaneous top-down and bottom-up nanofabrication approach called simultaneous plasma enhanced reactive ion synthesis and etching (SPERISE). For the first time the atomic addition and subtraction of nanomaterials are concurrently observed and precisely controlled in a single-step process permitting ultrahigh-throughput, lithography-less, wafer-scale, and room-temperature nanomanufacturing. Rapid low-cost manufacturing of high-density, high-uniformity, light-trapping nanocone arrays was demonstrated on single crystalline and polycrystalline silicon wafers, as well as amorphous silicon thin films. The proposed nanofabrication mechanisms also provide a general guideline to designing new SPERISE methods for other solid-state materials besides silicon.
Assuntos
Nanoestruturas , Nanotecnologia/métodos , Silício/química , Cristalização , Fontes de Energia Elétrica , Indústrias , Luz , Reprodutibilidade dos Testes , Energia Solar , Fatores de TempoRESUMO
We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.
Assuntos
Imageamento Tridimensional/métodos , Microscopia de Fluorescência/métodos , Nanopartículas/química , Animais , Células CHO , Sobrevivência Celular , Cricetinae , Cricetulus , Corantes Fluorescentes/metabolismo , Nanopartículas/ultraestruturaRESUMO
Molecular probe arrays printed on solid surfaces such as DNA, peptide, and protein microarrays are widely used in chemical and biomedical applications especially genomic and proteomic studies (Pollack et al 1999 Nat. Genet. 23 41-6, Houseman et al 2002 Nat. Biotechnol. 20 270-4, Sauer et al 2005 Nat. Rev. Genet. 6 465-76) as well as surface imaging and spectroscopy (Mori et al 2008 Anal. Biochem. 375 223-31, Liu et al 2006 Nat. Nanotechnol. 1 47-52, Liu 2010 IEEE J. Sel. Top. Quantum Electron. 16 662-71). Unfortunately the printed molecular spots on solid surfaces often suffer low distribution uniformity due to the lingering 'coffee stain' (Deegan et al 1997 Nature 389 827-9) problem of molecular accumulations and blotches, especially around the edge of deposition spots caused by solvent evaporation and convection processes. Here we present, without any surface chemistry modification, a unique solid surface of high-aspect-ratio silver-coated silicon nanocone arrays that allows highly uniform molecular deposition and thus subsequent uniform optical imaging and spectroscopic molecular detection. Both fluorescent Rhodamine dye molecules and unlabeled oligopeptides are printed on the metallic nanocone photonic substrate surface as circular spot arrays. In comparison with the printed results on ordinary glass slides and silver-coated glass slides, not only high printing density but uniform molecular distribution in every deposited spot is achieved. The high-uniformity and repeatability of molecular depositions on the 'coffee stain'-free nanocone surface is confirmed by laser scanning fluorescence imaging and surface enhanced Raman imaging experiments. The physical mechanism for the uniform molecular deposition is attributed to the superhydrophobicity and localized pinned liquid-solid-air interface on the silver-coated silicon nanocone surface. The unique surface properties of the presented nanocone surface enabled high-density, high-uniformity probe spotting beneficial for genomic and proteomic microarrays and surface molecular imaging.
Assuntos
Nanoestruturas/química , Nanotecnologia/métodos , Fenômenos Ópticos , Fótons , Prata/química , Análise Espectral Raman , Propriedades de SuperfícieRESUMO
Surface enhanced Raman spectroscopy (SERS) has been increasingly utilized as an analytical technique with significant chemical and biological applications (Qian et al 2008 Nat. Biotechnol. 26 83; Fujita et al 2009 J. Biomed. Opt. 14 024038; Chou et al 2008 Nano Lett.8 1729; Culha et al 2003 Anal. Chem. 75 6196; Willets K A 2009 Anal. Bioanal. Chem. 394 85; Han et al 2009 Anal. Bioanal. Chem. 394 1719; Sha et al 2008 J. Am. Chem. Soc. 130 17214). However, production of a robust, homogeneous and large-area SERS substrate with the same ultrahigh sensitivity and reproducibility still remains an important issue. Here, we describe a large-area ultrahigh-uniformity tapered silver nanopillar array made by laser interference lithography on the entire surface of a 6 inch wafer. Also presented is the rigorous optical characterization method of the tapered nanopillar substrate to accurately quantify the Raman enhancement factor, uniformity and repeatability. An average homogeneous enhancement factor of close to 10(8) was obtained for benzenethiol adsorbed on a silver-coated nanopillar substrate.