From functional nanostructured surfaces to innovative optical biosensors Giacomo Dacarro Dipartimento di Fisica A.Volta Dipartimento di Chimica Generale Università degli Studi di Pavia Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
utline Selfassembled monolayer (SAM) formation and growth Metal nanoparticles (NP) anchoring on SAM Metal ions and NP SAM as antibacterial materials Applications to photonic biosensors Plasmonicphotonic biosensor SENSe (surfaceenhanced nanosensor) Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
SAM formation and growth 1. Si 2 surface cleaning/activation 2. Silanization 3. Further growth of the SAM X X X X X X X H H H H Si Si Si Si Me Si Me Me H Si H H H Si H Si Si Si Si Si Si Si Si Si Si Si Si Si 2 surfaces: glass, quartz, Si 2 /Si ther surfaces: Si 3 N 4, inorganic oxides 3D Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
F F F F F molecular monolayer growth (via specific coupling reactions) formation of a nanoparticles monolayer (via specific XNP interactions) NP Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
SH chromophore Me Si Me Me N N = CH N Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Contact Angle cleaning ~45 <15 6065 surface hydrophilicity/lipophilicity silanization Spectroscopic Ellipsometry Surface dielectric function changes SAM thickness 1.71.8 nm Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
FTIRAttenuated Total Reflectance (ATR) CH C= Identification of chemical groups Atomic Force Microscopy Sample topography, morphology Roughness (rms roughness 0.31nm ) 0.25 µm Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Synthesis and anchoring of Ag nanoparticles Silver salt: AgN 3 Stabilizing agent: sodium citrate Reducing agent: NaBH 4 SPR peak NP radius 10nm Functionalized glass dipped in NP solution (rt, overnight) SH SH SH SH SH SH Si Si Si Si Si Si Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Stability of Ag NP monolayer 1 µm 0.25 µm [Ag] = 4.84 µg/cm 2 (Neutron Activation Analysis) SAM is stable in air, water, PBS Ag release in water (ICP) 1.1 % Time evolution of SPR peak intensity Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Anchoring of metal ion complexes Ligand Linker = + + + + + + + + + Silane NP bound through weak (non covalent) interactions FTIRATR UVVisible spectroscopy 0.0030 0.12 0.0025 Metal complex CT 0.10 SPR ATR signal Transmittance Abs / a.u. 0.0020 0.0015 Abs 0.08 0.06 ATR spectrum on SAM bulk FTIR spectrum of silanecomplex 0.0010 0.04 3500 3000 2500 wavenumber, cm 1 2000 1500 0.0005 200 300 400 500 600 700 800 Wavelength / nm 300 400 500 600 700 800 wavelength, nm Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Antibacterial activity Ag NP on SH SAM Cu complex SAM + + + + + + + + Cu complex SAM with Ag NP overlayer + + + + + + + + Strands Staphylococcus aureus (Gram+) Escherichia Coli (Gram) Strands Staphylococcus aureus (Gram+) Escherichia Coli (Gram) Strands Staphylococcus aureus (Gram+) Escherichia Coli (Gram) Microbicidal Effect ME = log N C log N E + = ME 5 h 24 h 1.22 5 3.24 4.12 ME 5 h 24 h 0.88 1.71 2.29 2.5 ME 5 h 24 h 2.33 6.60 5.29 7.06 Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
F F F F Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009 A tertiary monolayer HS NH 2 H H N H N H H NH 2 SH SH N H N H S
Photonic biosensors Nanofabrication Techniques to develop Sensitive Materials photonic or plasmonic structures, porous substrates, nanoparticle deposits, light emitters + Spectroscopic Nanooptical Techniques determine the optical response of the devices in presence of biomolecules PB possess sensing ability due to the high specificity of the optical response of the environment trough the ambient dielectric function ~ ε = ~ n 2 = (n + ik) 2, the absorption coefficient α or absorption cross section σ
Bloch Surface Waves (BSW) BSW can be considered as the dielectric analogues of SP ε1 σla,εa Lb,εb La,εa asin:h 1D PhC Both TE and TM polarizations The mode can be excited through a propagating wave Dispersion can be tailored by changing layer thicknesses In principle they can be free from losses M. Liscidini and J. E.Sipe, JSA b, 2009, 26, 279 Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Enhancement of emission by BSW A rhodamine monolayer is covalently attached to the surface of the multilayer. A strong modification of Rhodamine emission spectrum is observed, due to the excitation of BSW 532 nm pump M. Liscidini et al., APL, 2009, 94, 043117 M. Liscidini et al., ptics Letters, 2009, 34(15), 2318 Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Towards the nanoscale chemical mapping Microscale Delivery of energy to nanoscale: adiabatic conversion of propagating EM wave to local fields Enhancement and control of the local nanoscale e.m. fields. Enhanced nearfield responses AFM topography at nanoscale resolution 2 µm R=5n m Far field optical collection if possible Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
A single QD Raman spectrum Few molecules sensitivity Amine peak estimated 10 NH 2 groups (from company linkage data) Maximum 80 NH 2 groups, SERS enh. 10 5 F. De Angelis et al., Nano Letters, 2008, 8, 2321 Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
SENSe overall device Nanometric resolution on real samples AFM topography+ Raman Scattering signals AFM cantilever The dual beam @ BINEM lab FIB milling + ebeam induced deposition Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Conclusions and perspectives Starting point Solution and surface chemistry ptics and photonics Goals Smart, customizable, functional selfassembled materials High sensitivity optical biosensors Probes for nanoscale analysis and singlemolecule detection Perspective Material science can provide powerful tools to bring medicine (diagnostics, drug delivery, targeting) to the molecular level Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009
Thanks to NanoLab Dipartimento di Chimica Generale P. Pallavicini A.Taglietti Y. Diaz Fernandez L. Pasotti G. Santucci E. Amato D. Barbieri Laboratorio di spettroscopia ottica Dipartimento di Fisica A.Volta G. Guizzetti M. Patrini M. Galli M. Liscidini L. Cucca (Atomic absorption) L. Bergamaschi (NAA) Dipartimento di Farmacologia Sperimentale e Applicata C. Dacarro P. Grisoli Dalla scienza dei materiali alla biomedicina molecolare Pavia 17/11/2009