We present a handheld biosensor system for the label-free and specific multiplexed detection of several biomarkers employing a spectrometer-free imaging measurement system. A photonic crystal surface functionalized with multiple specific ligands forms the optical transducer. The photonic crystal slab is fabricated on a glass substrate by replicating a periodic grating master stamp with a period of 370 nm into a photoresist via nanoimprint lithography and deposition of a 70-nm titanium dioxide layer. Capture molecules are coupled covalently and drop-wise to the photonic crystal surface. With a simple camera and imaging optics the surface-normal transmission is detected. In the transmission spectrum guided-mode resonances are observed that shift due to protein binding. This shift is observed as an intensity change in the green color channel of the camera. Non-functionalized image sections are used for continuous elimination of background drift. In a first experiment we demonstrate the specific and time-resolved detection of 90.0 nm CD40 ligand antibody, 90.0 nM EGF antibody, and 500 nM streptavidin in parallel on one sensor chip. In a second experiment, aptamers with two different spacer lengths are used as receptor. The binding kinetics with association and dissociation of 250 nM thrombin and regeneration of the sensor surface with acidic tris-HCl-buffer (pH 5.0) is presented for two measurement cycles.
Photonic crystal technology has attracted large interest in the last years. The possibility to generate highly sensitive sensor elements with photonic crystal structures is very promising for medical or environmental applications. The low-cost fabrication on the mass scale is as advantageous as the compactness and reliability of photonic crystal biosensors. The possibility to integrate microfluidic channels together with photonic crystal structures allows for highly compact devices. This article reviews different types of photonic crystal sensors including 1D photonic crystal biosensors, biosensors with photonic crystal slabs, photonic crystal waveguide biosensors and biosensors with photonic crystal microcavities. Their applications in biomolecular and pathogen detection are highlighted. The sensitivities and the detection limits of the different biosensors are compared. The focus is on the possibilities to integrate photonic crystal biosensors on-chip.
There is a strong need for low-cost biosensors to enable rapid, on-site analysis of biological, biomedical, or chemical substances. We propose a platform for label-free optical biosensors based on applying the analyte onto a surface-functionalized photonic crystal slab and performing a transmission measurement with two crossed polarization filters. This dark-field approach allows for efficient background suppression as only the photonic crystal guided-mode resonances interacting with the functionalized surface experience significant polarization rotation. We present a compact biosensor demonstrator using a low-cost light emitting diode and a simple photodiode capable of detecting the binding kinetics of a 2.5 nM solution of the protein streptavidin on a biotin-functionalized photonic crystal surface.
We present an experimental method for direct analysis of guided-mode resonances in photonic crystal slab structures using transmission measurements. By positioning the photonic crystal slab between orthogonally oriented polarization filters light transmission is suppressed except for the guided-mode resonances. Angle resolved transmission measurements with crossed polarizers are performed to obtain the band structure around the Gamma-point. Results are compared to mode simulations. Spatially resolved measurements in a confocal microscope setup are used for homogeneity characterizations. Stitching errors and inhomogeneities in exposure dose down to 1.3% in photonic crystal slabs fabricated by electron beam lithography are observed using this method.
For label-free assays employing photonic crystal slabs (PCSs), the sensitivity is one of the most important properties influencing the detection limit. We investigate the bulk sensitivity and the surface sensitivity of 24 different PCSs fabricated by injection molding of PMMA and subsequent sputtering of a Ta 2 O 5 high-index layer. The duty cycle of the linear grating is varied in steps of 0.1 between 0.2 and 0.7. Four different Ta 2 O 5 layer thicknesses (89 nm, 99 nm, 189 nm, 301 nm) are deposited. Both bulk and surface sensitivity are optimal for a Ta 2 O 5 layer thickness of 99 nm. The maximum bulk sensitivity of 138 nm/RIU is achieved for a duty cycle of 0.7, while the maximum surface sensitivity of 47 nm/RIU is obtained for a duty cycle of 0.5. Good agreement between experimental results and finite-difference time-domain (FDTD) simulations is observed. The PCSs sensitivity is linked to the mode intensity distribution. Laing, "Label-free assays on the BIND system," J. Biomol. Screen. 9(6), 481-490 (2004). 9. Y. Nazirizadeh, U. Bog, S. Sekula, T. Mappes, U. Lemmer, and M. Gerken, "Low-cost label-free biosensors using photonic crystals embedded between crossed polarizers," Opt. Express 18(18), 19120-19128 (2010). 10. I. M. White and X. Fan, "On the performance quantification of resonant refractive index sensors," Opt. Express 16(2), 1020-1028 (2008). 11. M. El Beheiry, V. Liu, S. Fan, and O. Levi, "Sensitivity enhancement in photonic crystal slab biosensors," Opt.Express 18(22), 22702-22714 (2010). 12. U. Geyer, J. Hauss, B. Riedel, S. Gleiss, U. Lemmer, and M. Gerken, "Large-scale patterning of indium tin oxide electrodes for guided mode extraction from organic light-emitting diodes," J. Appl. Phys. 104(9), 093111 (2008). 13. L. J. Guo, "Nanoimprint lithography: methods and material requirements," Adv. Mater. 19(4), 495-513 (2007
We report a method enabling intensity-based readout for label-free cellular assays, and realize a reader device with the same footprint as a microtiter plate. For unambiguous resonance intensity measurements in resonance waveguide grating (RWG) sensors, we propose to apply resonances near the substrate cutoff wavelength. This method was validated in bulk refractive index, surface bilayer and G protein-coupled receptor (GPCR) experiments. The significantly reduced size of the reader device opens new opportunities for easy integration into incubators or liquid handling systems.
We propose and demonstrate a visual, all-optical pressure-measuring device composed of a flexible membrane dilating toward a photonic crystal slab. Due to its transparency and capability to be miniaturized, it may be integrated on the inner side of an artificial lens and directly measure the eye's intraocular pressure. Using crossed polarization filters for the readout process, we obtain a contrast enhancement for the circular contact area of the membrane with the photonic crystal slab. We demonstrate that the visible circle increases as a function of pressure.
Two methods for the fabrication of flexible and stretchable photonic crystal slabs are demonstrated and compared. In both cases a periodically nanostructured polydimethylsiloxane (PDMS) membrane is used as substrate. The first method is based on oblique-angle vapor deposition of SiO as a high refractive index material onto the nanostructured membrane. The deposition is made at an angle of 45° to the surface. The grooves of the nanostructure are aligned such that shading effects cause an inhomogeneous layer thickness distribution on the surface. This supports controlled, periodic cracking of the high index layer upon stretching. In the second approach ZnO nanoparticles are spin-coated on the nanostructured PDMS membrane. Here, the membrane can be stretched and serves as a photonic crystal slab without the need of any further treatment. For both types of flexible photonic crystal slabs a shift of the guided mode resonances to longer wavelengths is observed upon stretching. For a 20% strain perpendicular to the grating grooves a resonance shift of more than 50 nm is obtained.
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