Lensfree optofluidic plasmonic sensor for real-time and label-free monitoring of molecular binding events over a wide field-of-view

We demonstrate a high-throughput biosensing device that utilizes microfluidics based plasmonic microarrays incorporated with dual-color on-chip imaging toward real-time and label-free monitoring of biomolecular interactions over a wide field-of-view of >20 mm^2. Weighing 40 grams with 8.8 cm in height, this biosensor utilizes an opto-electronic imager chip to record the diffraction patterns of plasmonic nanoapertures embedded within microfluidic channels, enabling real-time analyte exchange. This plasmonic chip is simultaneously illuminated by two different light-emitting-diodes that are spectrally located at the right and left sides of the plasmonic resonance mode, yielding two different diffraction patterns for each nanoaperture array. Refractive index changes of the medium surrounding the near-field of the nanostructures, e.g., due to molecular binding events, induce a frequency shift in the plasmonic modes of the nanoaperture array, causing a signal enhancement in one of the diffraction patterns while suppressing the other. Based on ratiometric analysis of these diffraction images acquired at the detector-array, we demonstrate the proof-of-concept of this biosensor by monitoring in real-time biomolecular interactions of protein A/G with immunoglobulin G (IgG) antibody. For high-throughput on-chip fabrication of these biosensors, we also introduce a deep ultra-violet lithography technique to simultaneously pattern thousands of plasmonic arrays in a cost-effective manner

Field-portable optofluidic plasmonic biosensor for wide-field and label-free monitoring of molecular interactions

We demonstrate a field-portable optofluidic plasmonic sensing device, weighing 40 g and 7.5 cm in height, which merges plasmonic microarrays with dual-wavelength lensfree on-chip imaging for real-time monitoring of protein binding kinetics

Handheld high-throughput plasmonic biosensor using computational on-chip imaging

We demonstrate a handheld on-chip biosensing technology that employs plasmonic microarrays coupled with a lens-free computational imaging system towards multiplexed and high-throughput screening of biomolecular interactions for point-of-care applications and resource-limited settings. This lightweight and field-portable biosensing device, weighing 60 g and 7.5 cm tall, utilizes a compact optoelectronic sensor array to record the diffraction patterns of plasmonic nanostructures under uniform illumination by a single-light emitting diode tuned to the plasmonic mode of the nanoapertures. Employing a sensitive plasmonic array design that is combined with lens-free computational imaging, we demonstrate label-free and quantitative detection of biomolecules with a protein layer thickness down to 3 nm. Integrating large-scale plasmonic microarrays, our on-chip imaging platform enables simultaneous detection of protein mono- and bilayers on the same platform over a wide range of biomolecule concentrations. In this handheld device, we also employ an iterative phase retrieval-based image reconstruction method, which offers the ability to digitally image a highly multiplexed array of sensors on the same plasmonic chip, making this approach especially suitable for high-throughput diagnostic applications in field settings

A novel approach to locate Phytophthora infestans resistance genes on the potato genetic map

Mapping resistance genes is usually accomplished by phenotyping a segregating population for the resistance trait and genotyping it using a large number of markers. Most resistance genes are of the NBS-LRR type, of which an increasing number is sequenced. These genes and their analogs (RGAs) are often organized in clusters. Clusters tend to be rather homogenous, viz. containing genes that show high sequence similarity with each other. From many of these clusters the map position is known. In this study we present and test a novel method to quickly identify to which cluster a new resistance gene belongs and to produce markers that can be used for introgression breeding. We used NBS profiling to identify markers in bulked DNA samples prepared from resistant and susceptible genotypes of small segregating populations. Markers co-segregating with resistance can be tested on individual plants and directly used for breeding. To identify the resistance gene cluster a gene belongs to, the fragments were sequenced and the sequences analyzed using bioinformatics tools. Putative map positions arising from this analysis were validated using markers mapped in the segregating population. The versatility of the approach is demonstrated with a number of populations derived from wild Solanum species segregating for P. infestans resistance. Newly identified P. infestans resistance genes originating from S. verrucosum, S. schenckii, and S. capsicibaccatum could be mapped to potato chromosomes 6, 4, and 11, respectively

Actively transporting virus like analytes with optofluidics for rapid and ultrasensitive biodetection

Effective analyte delivery is essential to achieve rapid and sensitive biodetection systems. In this article, we present an actively controlled fluidic system integrated with a suspended plasmonic nanohole sensor to achieve superior analyte delivery efficiency and ultrafast sensor response, as compared to conventional fluidic systems. 70 nm sized virus like analyte solution is used to experimentally demonstrate the system performance improvements. Sensor response time is reduced by one order of magnitude as compared to the conventional methods. A seven orders of magnitude dynamic concentration range from 10(3) to 10(9) particles mL(-1) is quantified, corresponding to a concentration window relevant to clinical diagnosis and drug screening. Our non-destructive detection system, by enabling efficient analyte delivery, fast sensing response and minimal sample volume, opens up opportunities for sensitive, rapid and real-time virus detection in infectious disease control and point-of-care applications

Reusable Nanostencils for Creating Multiple Biofunctional Molecular Nanopatterns on Polymer Substrate

In this paper, we demonstrate a novel method for high throughput patterning of bioprobes with nanoscale features on biocompatible polymer substrate. Our technique, based on nanostencil lithography, employs high resolution and robust masks integrated with array of reservoirs. We show that the smallest pattern size can reach down to 100 nm. We also show that different types of biomolecules can be patterned on the same substrate simultaneously. Furthermore, the stencil can be reused multiple times to generate a series of identical patterns at low cost. Finally, we demonstrate that biomolecules can be covalently patterned on the surface while retaining their biofunctionalities. By offering the flexibility on the nanopattern design and enabling the reusability of the stencil, our approach significantly simplifies the bionanopatterning process and therefore could have profound implications in diverse biological and medical applications

Fabrication of Sub-10-nm Plasmonic Gaps for Ultra-Sensitive Raman Spectroscopy

The past two decades have witnessed the explosion of activities in the field of surface enhanced Raman spectroscopy (SERS). SERS platforms employ nano-structures that excite plasmonic modes with large local electromagnetic fields localized within small gap spaces between each constituting feature. Although the research-oriented SERS platforms yield significant signal enhancements to identify even single molecules, practical SERS-based sensors have not been fully introduced yet. The main reason behind this absence is the need for a cost-effective and reliable manufacturing method for controllable fabrication of plasmonic nano-gaps over large areas. In this article, we introduced a novel manufacturing process that enables fast and scalable fabrication of highly uniform sub-10-nm gaps that could yield large SERS signals. In this process, a conventional electroplating technique is used to produce unique nano-mushroom antenna arrays on a conducting substrate, resulting in controllable gap spaces between mushroom heads. By understanding the nature of mushroom shape antenna formation, we demonstrated the control of inter-metallic gaps down to 5 nm. We showed that the manufactured nano-structures yield Raman enhancements more than 10(8). Providing such large SERS signals that are uniform over large areas, our cost-effective fabrication technique could be very critical to realize practical SERS devices

Plasmonic Nanohole Arrays on a Robust Hybrid Substrate for Highly Sensitive Label-Free Biosensing

Plasmonic nanohole arrays have received significant attention, as they have highly advantageous optical properties for ultrasensitive and label-free biosensing applications. Currently, most of these subwavelength periodic apertures are mainly implemented on transparent materials, which results in multiple spectrally close transmission resonances. However, this spectral characteristic is not ideal for biosensing applications, as it complicates monitoring spectral variations. In this article, utilizing a hybrid substrate composed of a high refractive index dielectric interlayer over a transparent material, we show that gold nanohole arrays support spectrally isolated and well-defined plasmonic resonances that are easy to track. Compared to conventional configurations on transparent material, nanoholes on a hybrid substrate also exhibit plasmonic modes with well-preserved amplitudes, which is useful for reliable spectral monitoring. We show that nanohole arrays on a hybrid substrate are more sensitive to changes in surface conditions. Using a spectral integration method, which evaluates wavelength shifts in a large spectral window instead of monitoring only the plasmonic resonance wavelength, we obtain a detection limit as low as 2 x 10^(-5) RIU. Furthermore, we successfully demonstrate real-time monitoring of biomolecular binding interactions even at sub-1 ng/mL levels

Hand-Held Plasmonic Biosensor for High-Throughput Sensing for Point-of-Care Applications

We introduce a hand-held biosensor based on large-area plasmonic microarrays coupled with a lens-free computational imaging system for high-throughput biosensing. Our light-weight device, 60 g and 7.5 cm, is highly suitable for point-of-care applications for field-settings