Digital microarrays: single-molecule readout with interferometric detection of plasmonic nanorod labels

DNA and protein microarrays are a high-throughput technology that allow the simultaneous quantification of tens of thousands of different biomolecular species. The mediocre sensitivity and limited dynamic range of traditional fluorescence microarrays compared to other detection techniques have been the technology’s Achilles’ heel and prevented their adoption for many biomedical and clinical diagnostic applications. Previous work to enhance the sensitivity of microarray readout to the single-molecule (“digital”) regime have either required signal amplifying chemistry or sacrificed throughput, nixing the platform’s primary advantages. Here, we report the development of a digital microarray which extends both the sensitivity and dynamic range of microarrays by about 3 orders of magnitude. This technique uses functionalized gold nanorods as single-molecule labels and an interferometric scanner which can rapidly enumerate individual nanorods by imaging them with a 10× objective lens. This approach does not require any chemical signal enhancement such as silver deposition and scans arrays with a throughput similar to commercial fluorescence scanners. By combining single-nanoparticle enumeration and ensemble measurements of spots when the particles are very dense, this system achieves a dynamic range of about 6 orders of magnitude directly from a single scan. As a proof-of-concept digital protein microarray assay, we demonstrated detection of hepatitis B virus surface antigen in buffer with a limit of detection of 3.2 pg/mL. More broadly, the technique’s simplicity and high-throughput nature make digital microarrays a flexible platform technology with a wide range of potential applications in biomedical research and clinical diagnostics.The authors wish to thank Oguzhan Avci and Jacob Trueb for thoughtful comments and suggestions regarding numerical optimization of the optical system. This work was funded in part by a research contract with ASELSAN, Inc. and the Wallace H. Coulter Foundation 2010 Coulter Translational Award. (ASELSAN, Inc.; Wallace H. Coulter Foundation Coulter Translational Award)Accepted manuscrip

Chirality dependence of the radial breathing phonon mode density in single wall carbon nanotubes

A mass and spring model is used to calculate the phonon mode dispersion for single wall carbon nanotubes (SWNTs) of arbitrary chirality. The calculated dispersions are used to determine the chirality dependence of the radial breathing phonon mode (RBM) density. Van Hove singularities, usually discussed in the context of the single particle electronic excitation spectrum, are found in the RBM density of states with distinct qualitative differences for zig zag, armchair and chiral SWNTs. The influence the phonon mode density has on the two phonon resonant Raman scattering cross-section is discussed.Comment: 6 pages, 2 figures, submitted to Phys. Rev.

Tunable Resonant Raman Scattering from Singly Resonant Single Wall Carbon Nanotubes

We perform tunable resonant Raman scattering on 17 semiconducting and 7 metallic singly resonant single wall carbon nanotubes. The measured scattering cross-section as a function laser energy provides information about a tube's electronic structure, the lifetime of intermediate states involved in the scattering process and also energies of zone center optical phonons. Recording the scattered Raman signal as a function of tube location in the microscope focal plane allows us to construct two-dimensional spatial maps of singly resonant tubes. We also describe a spectral nanoscale artifact we have coined the "nano-slit effect"

Digital detection of exosomes by interferometric imaging

Exosomes, which are membranous nanovesicles, are actively released by cells and have been attributed to roles in cell-cell communication, cancer metastasis, and early disease diagnostics. The small size (30–100 nm) along with low refractive index contrast of exosomes makes direct characterization and phenotypical classification very difficult. In this work we present a method based on Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows multiplexed phenotyping and digital counting of various populations of individual exosomes (>50 nm) captured on a microarray-based solid phase chip. We demonstrate these characterization concepts using purified exosomes from a HEK 293 cell culture. As a demonstration of clinical utility, we characterize exosomes directly from human cerebrospinal fluid (hCSF). Our interferometric imaging method could capture, from a very small hCSF volume (20 uL), nanoparticles that have a size compatible with exosomes, using antibodies directed against tetraspanins. With this unprecedented capability, we foresee revolutionary implications in the clinical field with improvements in diagnosis and stratification of patients affected by different disorders.This work was supported by Regione Lombardia and Fondazione Cariplo through POR-FESR, project MINER (ID 46875467); Italian Ministry of Health, Ricerca Corrente. This work was partially supported by The Scientific and Technological Research Council of Turkey (grant #113E643). (Regione Lombardia; 46875467 - Fondazione Cariplo through POR-FESR, project MINER; Italian Ministry of Health, Ricerca Corrente; 113E643 - Scientific and Technological Research Council of Turkey)Published versio

High-Speed High Effiency Large Area Resonant Cavity Enhanced p-I-n Photodiodes for Multimode Fiber Communications

Cataloged from PDF version of article.In this letter, we report AlGaAs–GaAs p-i-n photodiodes with a 3-dB bandwidth in excess of 10 GHz for devices as large as 60- m diameter. Resonant cavity enhanced photodetection is employed to improve quantum efficiency, resulting in more than 90% peak quantum efficiency at 850 nm

Robust Entanglement in Atomic Systems via Lambda-Type Processes

It is shown that the system of two three-level atoms in $\Lambda$ configuration in a cavity can evolve to a long-lived maximum entangled state if the Stokes photons vanish from the cavity by means of either leakage or damping. The difference in evolution picture corresponding to the general model and effective model with two-photon process in two-level system is discussed.Comment: 10 pages, 3 figure

Deformable Model for 3D Intramodal Nonrigid Breast Image Registration with Fiducial Skin Markers

We implemented a new approach to intramodal non-rigid 3D breast image registration. Our method uses fiducial skin markers (FSM) placed on the breast surface. After determining the displacements of FSM, finite element method (FEM) is used to distribute the markers’ displacements linearly over the entire breast volume using the analogy between the orthogonal components of the displacement field and a steady state heat transfer (SSHT). It is valid because the displacement field in x, y and z direction and a SSHT problem can both be modeled using LaPlace’s equation and the displacements are analogous to temperature differences in SSHT. It can be solved via standard heat conduction FEM software with arbitrary conductivity of surface elements significantly higher than that of volume elements. After determining the displacements of the mesh nodes over the entire breast volume, moving breast volume is registered to target breast volume using an image warping algorithm. Very good quality of the registration was obtained. Following similarity measurements were estimated: Normalized Mutual Information (NMI), Normalized Correlation Coefficient (NCC) and Sum of Absolute Valued Differences (SAVD). We also compared our method with rigid registration technique