Exploring metallic nanoparticles in Biophotonics
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2 Exploring metallic nanoparticles in Biophotonics Renato E. de Araujo Laboratório de Óptica Biomédica e Imagem, Universidade Federal de Pernambuco Recife, PE, Brazil. renato.earaujo@ufpe.br
3 LABORATÓRIO DE ÓPTICA BIOMÉDICA E IMAGEM 1) Spectroscopy 2) Microscopy Normalized Fluorescence (a.u.) Growth Medium Fusarium T. Schoenleinii T. Rubrum Wavelength (nm) spectrometer 3) Dengue 4) OCT (Ganymed Thorlabas) 5) Metallic nanoparticles...
4 Outline Light -NANO particle interaction Metal Enhanced Fluorescence Biosensors Photodynamic therapy Imaging technique Conclusion spheres rods shells cages
5 Light -NANO particle interaction x h m h in n E E ˆ = ε ε ε ) ˆ sin ˆ (2cos θ θ θ ε ε ε ε n n r a E E r h m h m ext = Bohren CF, Huffman DR. (1983) Absorption and Scattering of Light by Small Particles
6 Resonant enhancement condition: Light -NANO particle interaction ( h ε NP + 2ε ) Minimum Localized Plasmon Resonance ε 2 = in NP εh E and E ext increase Kelly KL, et al J Phys Chem B
7 Light -NANO particle interaction ε NP= Silver ε ( D, ω) Fluorophore -NANO particle interaction F
8 Fluorophore -NANO particle interaction Signal amplification mechanism of NPs in fluorescent biosensor. Metal Enhanced Fluorescence
9 Metal Enhanced Fluorescence F Energy transfer (FRET) ) ˆ sin ˆ (2cos θ θ θ ε ε ε ε n n r a E E r h m h m ext = ) / ( R r efficiency + = Dulkeith, Phys. Rev. Lett. 2002
10 MEF: Great potential for advances in medical diagnostics and biotechnological methodology Film platform: Solution-base platform:
11 Metal Enhanced Fluorescence: Silver Nanospheres Normalized Emission Tryptophan f =0.45x10-6 f =1.5x10-6 f =2.4x10-6 f =4.35x10-6 F Distance is important Wavelength (nm)
12 Metal Enhanced Fluorescence: Silver Nanospheres Normalized Emission Tryptophan f =0.45x10-6 f =1.5x10-6 f =2.4x10-6 f =4.4x Wavelength (nm) Enhanced Multi-Photon Excitation of Tryptophan- Silver Colloid Renato E. de Araujo, Diego Rativa and Anderson S. L. Gomes Metal-Enhanced Fluorescence. Edited by Chris D. Geddes. Copyright 2010 John Wiley & Sons, Inc. ISBN-10: ISBN-13: John Wiley & Son
13 Mettal nanoparticle in biomedical applications Mettal nanoparticle based biosensors Growth of In Vitro Diagnostic Market US$ 43 Billion in 2010 US$ 52 Billion by 2013-end. Electrochemical biosensors Amplified voltammetric detection (imobilization plataform, Immunoassay), Capillary and gel electrophoresis. Optical biosensor Glucose biosensor (SERS) ph-sensor (SERS) Pregnancy test (colormetric response) Detection of Biomolecules (DNA, Enzymes...) Jin R, et al (2003) J Am Chem Soc
14 Why explore Quantum Dots? Quantum Dots Biosensors Quantum dots (QDs) - nanocrystalline semiconductors CdTe/CdS Evident Technologies Inc. CdTe/CdS core-shell quantum dots have an inner spherical CdTe semiconductor crystal surrounded by a shell of CdS semiconductor. Multiplexed Diagnostics Quantum Yield 20-30%
15 Mettal nanoparticle in biomedical applications QD-based Biosensors ph Probes Ion Sensors (Zn2+, Cu2+, Hg2+, Ag+, Fe3+, I-) Detection of Organic Compounds (Pesticides) Detection of Biomolecules (DNA, Proteins and Enzymes, amino acids) FRET fluorescence quenching (QDs are poor acceptors) Biosensors Specificity and Sensitivity
16 Qd Ag NP based Biosensors + =? Silver NP Qd Enhance Qd Fluorescence Enhance Biosensor Sensitivity
17 Qd Ag NP based Biosensors Materials - Silver NP Pectin Ag Ag + Pectin Gelification food industry
18 Materials - Silver NP Synthesis and characterization of silver nanoparticles Nps/mL
19 Materials - Silver NP Synthesis and characterization of silver nanoparticles Absorbance (a. u.) 1,0 0,8 0,6 0,4 Ag with Pectin Ag without Pectin Small shift ( 4 nm) of peak band indicates the presence of the pectin 0,2 0, W avelength(nm) Average diameter: 13 ± 5 nm
20 Qd Ag NP based Biosensors Ag + =? CdTe/CdS Ag + Pectin 0,6 1,2 Absorbance (a.u.) 0,5 0,4 0,3 0,2 0,1 1,0 0,8 0,6 0,4 0,2 Intensity (a.u.) Gelification food industry ,0 Wavelength (nm) 0,0
21 Qd Ag NP based Biosensors Intensity (a. u.) CdTe/CdS H2O CdTe/CdS Ag Pectin CdTe/CdS Ag Bared Wavelength (nm) Metal Enhanced Fluorescence: 100% increase of the CdTe/CdS emission
22 Mettal nanoparticle in biomedical applications Cancer therapy Photothermal therapy (Gold nanoshells and nanorods) Radiotherapy treatments (Increased absorption of ionizing radiation by the gold nanoparticles) Photodynamic therapy (Vehicle for the delivery of photosensitizers) J. T. Lin, et al. Proc. SPIE 7574, 2010 PDT - bacterial and fungal infection therapies
23 NP based PDT photosensitizer + =? Ag Riboflavin 1,0 Riboflavin Ag with Pectin Ag without Pectin 1,0 0,8 0,8 Absorbance (a. u.) 0,6 0,4 0,6 0,4 0,2 0,2 0,0 0, Wavelength(nm) Metal Enhanced Singlet Oxygen Generation
24 Metal Enhanced Singlet Oxygen Generation: Ag + =? Spectral properties of Singlet Oxygen Sensor Green (SOSG) before and after reaction with 1O2. Absorption (a, c) and fluorescence emission (b, d) spectra of SOSG before (a, b) and after (c, d) reaction with 1O2 (Flors e col, 2006).
25 NP based PDT photosensitizer 415 nm nm Pectin was used to improve metal enhanced singlet oxygen generation (28.5%)
26 Mettal nanoparticle in biomedical applications Cell and tissue imaging Scanning electron microscopy Optical microscopy (SERS, confocal, two-photon) Optical coherence tomography (Durr et al 2007)
27 In Situ Gold Nanoparticles formation: Contrast Agent for Dental Optical Coherence Tomography Contemporary restorative procedures are based on the adhesive properties of tooth colored resin-based materials Stable bonding is created by the fully impregnation of the dentin substrate by bleds of resin monomers, thus avoiding different degrees of incomplete impregnation and producing a compact and homogenous Hybrid layer.
28 In Situ Gold Nanoparticles formation: Contrast Agent for Dental Optical Coherence Tomography
29 Mettal nanoparticle in Dental Optical Coherence Tomography
30 Optical Coherence Tomography Spectral Domain OCT Equipment Resolution Stratus (Carl Zeiss) 10µm C7-XR OCT (St. Jude Medical) 15µm GANYMEDE SD-OCT System (Thorlabs) 5µm
31 Light source Gordan, Nature 2002 Solar collector (Daniel Farkas)
32 Solar OCT Light source spectrometer Resolution: 1.67 ± 0.02µm Variations of less than 0.2% Power: 1 mw
33 Conclusion
34
35 Laboratório de Óptica Biomédica e Imagem, Universidade Federal de Pernambuco Recife, PE, Brazil. POSITIONS AVAILABLE
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