-
[show abstract]
[hide abstract]
ABSTRACT: A laser line-scanning instrument was developed to optimize the near-field enhancement capability of a one-dimensional photonic crystal (PC) for excitation of surface-bound fluorophores. The excitation laser beam is shaped into an 8 μm × 1 mm line that is focused along the direction of the PC grating, while remaining collimated perpendicular to the grating. Such a beam configuration offers high excitation power density while simultaneously providing high resonant coupling efficiency from the laser to the PC surface. Using a panel of 21 immunofluorescence assays on the PC surface in a microarray format, the approach achieves an enhancement factor as high as 90-fold between on-resonance and off-resonance illumination. The instrument provides a capability for sensitive and inexpensive analysis of cancer biomarkers in clinical applications.
Optics Letters 07/2012; 37(13):2565-7. · 3.40 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A photonic crystal (PC) surface is demonstrated as a high-sensitivity platform for detection of a panel of 21 cancer biomarker antigens using a sandwich enzyme-linked immunosorbent assay (ELISA) microarray format. A quartz-based PC structure fabricated by nanoimprint lithography, selected for its low autofluorescence, supports two independent optical resonances that simultaneously enable enhancement of fluorescence detection of biomarkers and label-free quantification of the density of antibody capture spots. A detection instrument is demonstrated that supports fluorescence and label-free imaging modalities, with the ability to optimize the fluorescence enhancement factor on a pixel-by-pixel basis throughout the microarray using an angle-scanning approach for the excitation laser that automatically compensates for variability in surface chemistry density and capture spot density. Measurements show that the angle-scanning illumination approach reduces the coefficient of variation of replicate assays by 20-99% compared to ordinary fluorescence microscopy, thus supporting reduction in limits of detectable biomarker concentration. Using the PC resonance, biomarkers in mixed samples were detectable at the lowest concentrations tested (2.1-41 pg/mL), resulting in a three-log range of quantitative detection.
Analytical Chemistry 12/2011; 84(2):1126-33. · 5.86 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: By combining photonic crystal label-free biosensor imaging with photonic crystal enhanced fluorescence, it is possible to selectively enhance the fluorescence emission from regions of the PC surface based upon the density of immobilized capture molecules. A label-free image of the capture molecules enables determination of optimal coupling conditions of the laser used for fluorescence imaging of the photonic crystal surface on a pixel-by-pixel basis, allowing maximization of fluorescence enhancement factor from regions incorporating a biomolecule capture spot and minimization of background autofluorescence from areas between capture spots. This capability significantly improves the contrast of enhanced fluorescent images, and when applied to an antibody protein microarray, provides a substantial advantage over conventional fluorescence microscopy. Using the new approach, we demonstrate detection limits as low as 0.97 pg/ml for a representative protein biomarker in buffer.
Optics Express 11/2011; 19(23):23327-40. · 3.59 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report on the use of photonic crystal surfaces as a high-sensitivity platform for detection of a panel of cancer biomarkers in a protein microarray format. The photonic crystal surface is designed to provide an optical resonance at the excitation wavelength of cyanine-5 (Cy5), thus providing an increase in fluorescent intensity for Cy5-labeled analytes measured with a confocal microarray scanner, compared to a glass surface. The sandwich enzyme-linked immunosorbent assay (ELISA) is undertaken on a microarray platform to undertake a simultaneous, multiplex analysis of 24 antigens on a single chip. Our results show that the resonant excitation effect increases the signal-to-noise ratio by 3.8- to 6.6-fold, resulting in a decrease in detection limits of 6-89%, with the exact enhancement dependent upon the antibody-antigen interaction. Dose-response characterization of the photonic crystal antibody microarrays shows the capability to detect common cancer biomarkers in the <2 pg/mL concentration range within a mixed sample.
Analytical Chemistry 02/2011; 83(4):1425-30. · 5.86 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A Photonic Crystal (PC) surface fabricated upon a quartz substrate using nanoimprint lithography has been demonstrated to enhance light emission from fluorescent molecules in close proximity to the PC surface. Quartz was selected for its low autofluorescence characteristics compared to polymer-based PCs, improving the detection sensitivity and signal-to-noise ratio (SNR) of PC Enhanced Fluorescence (PCEF). Nanoimprint lithography enables economical fabrication of the subwavelength PCEF surface structure over entire 1x3 in2 quartz slides. The demonstrated PCEF surface supports a transverse magnetic (TM) resonant mode at a wavelength of λ = 632.8 nm and an incident angle of θ = 11°, which amplifies the electric field magnitude experienced by surface-bound fluorophores. Meanwhile, another TM mode at a wavelength of λ = 690 nm and incident angle of θ = 0° efficiently directs the fluorescent emission toward the detection optics. An enhancement factor as high as 7500 × was achieved for the detection of LD-700 dye spin-coated upon the PC, compared to detecting the same material on an unpatterned glass surface. The detection of spotted Alexa-647 labeled polypeptide on the PC exhibits a 330 × SNR improvement. Using dose-response characterization of deposited fluorophore-tagged protein spots, the PCEF surface demonstrated a 140 × lower limit of detection compared to a conventional glass substrate.
Optics Express 11/2010; 18(24):24793-808. · 3.59 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The effect of resonant fluorescent enhancement from a photonic crystal surface upon the fluorescent photobleaching rate of Cyanine-5 labeled protein has been investigated. We show that the enhanced excitation mechanism for photonic crystal enhanced fluorescence, in which the device surface resonantly couples light from an excitation laser, accelerates photobleaching in proportion to the coupling efficiency of the laser to the photonic crystal. We also show that the enhanced extraction mechanism, in which the photonic crystal directs emitted photons approximately normal to the surface, does not play a role in the rate of photobleaching. We show that the photobleaching rate of dye molecules on the photonic crystal surface is accelerated by 30x compared to an ordinary glass surface, but substantial signal gain is still evident, even after extended periods of continuous illumination at the resonant condition.
Journal of Fluorescence 11/2010; 21(2):707-14. · 2.11 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: This paper describes a novel fabrication technique for constructing a polymer-based large-core single-mode rib waveguide. A negative tone SU8 photoresist with a high optical transmission over a large wavelength range and stable mechanical properties was used as a waveguide material. A waveguide was constructed by using a polydimethylsiloxane stamp combined with a solvent-assisted microcontact molding technique. The effects on the final pattern's geometry of four different process conditions were investigated. Optical simulations were performed using beam propagation method software. Single-mode beam propagation was observed at the output of the simulated waveguide as well as the actual waveguide through the microscope image.
Applied Optics 10/2008; 47(25):4540-7. · 1.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A flexible high-resolution sensor capable of measuring the distribution of pressure beneath the foot via a microfabricated optical waveguide system is presented. The uniqueness of the system is in its batch fabrication process, which involves a microfabrication molding technique with polydimethylsiloxane (PDMS) as the optical medium. The sensor manufacturing technique is described in detail, the optical performance of the waveguides is quantified and the effect of using a matching fluid to improve fiber-coupling efficiency is demonstrated. Mechanical loading tests were performed on a 4 x 4 array with a 2-mm spacing between sensing elements. Loading displacement curves were obtained using a 0 to 0.4 mm triangle loading profile. A force of 0.28 N applied to one of the sensing elements produced a displacement of a 0.325 mm and 39% change in the output light intensity. Multiple loadings were conducted to demonstrate the repeatability of the sensor. A force image algorithm with a two-layer neural network system was used to identify four load magnitudes and four different shaped applicators. All four shapes were successfully identified with the neural network.
IEEE transactions on bio-medical engineering 03/2008; 55(2 Pt 1):614-25. · 2.15 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The authors report a polymeric based rib waveguide with U shape self-align fiber couplers system using a simple micromolding process with SU8 as a molding material and polydimethysiloxane as a waveguide material. The material is used for its good optical transparency, low surface tension, biocompatibility, and durability. Furthermore, the material is highly formable. This unique fabrication molding technique provides a means of keeping the material and manufacturing costs to a minimum. The self-align fiber couplers system also proves a fast and simple means of light coupling. The flexible nature of the waveguide material makes this process ideal for a potential wearable optical sensor.
Journal of vacuum science & technology. B, Microelectronics and nanometer structures: processing, measurement, and phenomena: an official journal of the American Vacuum Society 01/2008; 26(1):L13-L18. · 1.34 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We present SU-8 as an optical waveguide and its integration into a functional one- and two-dimensional (1-D and 2-D) scanning device. The waveguide is fabricated as a tapered structure and cantilever beam (100 times 85 times 2100 mum). It is mounted onto a 2-D coupled bimorph piezoelectric actuator and driven at resonant frequencies of 4.71 kHz in the vertical and 25 Hz in the horizontal. This raster motion allows for 2-D image scanning when in combination with a light source and detector. An analysis of the polymer's optical capabilities has shown mode coupling efficiencies of 96% and measured a total power output intensity of 20% with a 633-nm light source. The output power can be attributed to the butt-end coupling misalignment, scattering losses due to surface roughness, and absorption. The total vertical tip displacement is 669 mum with an angle of 25deg when driven with a 47-mum base excitation. A specific application for the design is in the area of endoscopy, where there is a need for a minimally invasive device that reduces the discomfort experienced by the patient. SU-8 as the functional waveguide is the initial phase in creating a microfabricated endoscope system.
Journal of Lightwave Technology 04/2007; 25(3):850-860. · 2.78 Impact Factor
