Point-of-Need bioanalytics based on planar optical interferometry.

This review brings about a comprehensive presentation of the research on interferometric transducers, which have emerged as extremely promising candidates for viable, truly-marketable solutions for PoN applications due to the attested performance that has reached down to 10(-8) in term of effective refractive index changes. The review explores the operation of the various interferometric architectures along with their design, fabrication, and analytical performance aspects. The issues of biosensor functionalization and immobilization of receptors are also addressed. As a conclusion, the comparison among them is attempted in order to delve into and acknowledge their current limitations, and define the future trends.

Monolithically integrated broad-band Mach-Zehnder interferometers for highly sensitive label-free detection of biomolecules through dual polarization optics.

Protein detection and characterization based on Broad-band Mach-Zehnder Interferometry is analytically outlined and demonstrated through a monolithic silicon microphotonic transducer. Arrays of silicon light emitting diodes and monomodal silicon nitride waveguides forming Mach-Zehnder interferometers were integrated on a silicon chip. Broad-band light enters the interferometers and exits sinusoidally modulated with two distinct spectral frequencies characteristic of the two polarizations. Deconvolution in the Fourier transform domain makes possible the separation of the two polarizations and the simultaneous monitoring of the TE and the TM signals. The dual polarization analysis over a broad spectral band makes possible the refractive index calculation of the binding adlayers as well as the distinction of effective medium changes into cover medium or adlayer ones. At the same time, multi-analyte detection at concentrations in the pM range is demonstrated.

All-silicon monolithic Mach-Zehnder interferometer as a refractive index and bio-chemical sensor.

A complete Mach-Zehnder interferometer monolithically integrated on silicon is presented and employed as a refractive index and bio-chemical sensor. The device consists of broad-band light sources optically coupled to photodetectors through monomodal waveguides forming arrays of Mach-Zehnder interferometers, all components being monolithically integrated on silicon through mainstream silicon technology. The interferometer is photonically engineered in a way that the phase difference of light travelling through the sensing and reference arms is approximately wavelength independent. Consequently, upon effective medium changes, it becomes feasible even with a broad-band source to induce sinusoidal-type of detector photocurrents similar to the classical monochromatic counterparts. The device is completed with its fluidic and interconnect components so that on chip interferometric measurements can be performed. Examples of refractive index and protein sensing are presented to establish the potential of the proposed device for real-time in situ monitoring applications. This is the only silicon device that has achieved complete on-chip interferometry.

Broad-band Mach-Zehnder interferometers as high performance refractive index sensors: theory and monolithic implementation.

Broad-band Mach-Zehnder interferometry is analytically described and experimentally demonstrated as an analytical tool capable of high accuracy refractive index measurements over a wide spectral range. Suitable photonic engineering of the interferometer sensing and reference waveguides result in sinusoidal TE and TM spectra with substantially different eigen-frequencies. This allows for the instantaneous deconvolution of multiplexed polarizations and enables large spectral shifts and noise reduction through filtering in the Fourier Transform domain. Due to enhanced sensitivity, optical systems can be designed that employ portable spectrum analyzers with nm range resolution without compromising the sensor analytical capability. Practical detection limits in the 10(-6)-10(-7) RIU range are achievable, including temperature effects. Finally, a proof of concept device is realized on a silicon microphotonic chip that monolithically integrates broad-band light sources and single mode silicon nitride waveguides. Refractive index detection limits rivaling that of ring resonators with externally coupled laser sources are demonstrated. Sensitivities of 20 µm/RIU and spectral shifts in the tens of a pm are obtained.

Molecular film growth monitoring via reflection microscopy on periodically patterned substrates.

An optical method is presented for in the situ monitoring of biomolecular films via reflection microscopy on patterned substrates. The method is based on measuring the reflection coefficient of a composite consisting of a substrate, a patterned optical layer, the thin film to be monitored and the cover medium. The optical layer is patterned so that an array of squares is surrounded by the bare substrate. The reflectance difference between the optical layer squares and the bare substrate is the observable, whose fractional changes reveal the thickness of the film through a simple analytical expression. The periodic image is recorded by a digital microscope, and through Fourier transform techniques, the normalized differential reflectance of the patterned optical composite is calculated as the contrast factor of two dimensional bit map. The method is demonstrated by measuring a protein binding assay inside a microfluidic module placed under a microscope.

Competitive technology approaches for electronic hybridization detection in a microsystem with microfluidics for diagnosis genetic tests.

This paper is presenting competitive technology alternatives for the electronic hybridization detection in a microsystem with microfluidics for diagnosis genetic tests that are carried out by two competitive research projects. The technologies developed are a photosensor, a capacitive sensor and an optical real-time affinity biosensor. The performance of those biosensors will be evaluated but also their manufacturability and cost will define the appropriateness of each one for industrialization and their integration on a microsystem for diagnosis genetic testing.

Multi-analyte capillary immunosensor for the determination of hormones in human serum samples.

In this work we present the development of a multi-analyte immunosensor for the determination of follitropin, human chorionic gonadotropin and prolactin in human serum. The immunosensor is based on plastic capillaries. According to the methodology, discrete areas of the internal capillary surface are coated with different antibodies, which are highly specific for each one of the analytes to be determined. The sample that will be analyzed along with a mixture of analyte-specific biotinylated antibodies is introduced into the capillary. The coated and the detection antibodies react with different epitopes of the analytes in the sample to form a 'sandwich'. The detection is based on reaction of the immobilized biotinylated antibody with streptavidin labeled with R-phycoerythrin. The fluorescent areas formed were quantified by scanning the capillary with a light beam of appropriate wavelength. A light sensor placed at the end of the capillary detects the emitted photons, that are trapped and waveguided into the capillary walls. The multi-analyte immunosensor assays were characterized by high specificity and short analysis time. In addition, the results obtained by the multi-analyte optical capillary immunosensor were comparable to those obtained by immunofluorimetric assays performed in microtitration wells. Potential applications of the proposed immunosensor include determination of several analyte panels in a broad spectrum of disciplines such as endocrinology, hematology, and oncology.

Electron beam patterning of biomolecules.

The patterning of biomolecules on semiconducting surfaces is of central importance in the fabrication of novel biodevices. In the process of patterning, it is required that the biomolecule preserves its properties and the substrate is not damaged by the chemicals, the temperatures or the patterning beams involved in the procedure. Recently, both DUV and electron beam microlithography have been used in order to deposit protein layers in predefined patterns. Various approaches have been used, some involving photoresists. Contrast between exposed and unexposed regions, resolution of adjacent features and sensitivity to dose variation, are the key issues. The approach followed in this paper consists of a direct patterning of a biotin layer, deposited on an amino-silane primed silicon nitride surface, using an electron beam. After irradiation, the surface is covered by bovine serum albumin (BSA), which acts as a blocking material to protect the exposed areas from streptavidin adsorption. Using 20 keV e-beam energy and doses, in the range 100-1000 microC/cm(2), submicrometer dense lines of 1-microm pitch have been obtained. The results have been tested by fluorescence optical microscopy.

A multi-band capillary immunosensor.

In the present work we propose a new optical immunosensor based on capillary geometry and capable of multianalyte determinations. The device is made of a polystyrene capillary tube. The inner walls of the capillary are segmented into distinct bands which are coated with appropriate binding molecules. Following excitation, some of the fluorescent photons emitted by the label are trapped and waveguided into the capillary walls provided they are launched towards the walls and within the critical angle. Here, Europium-labeled streptavidin reacted with different amounts of biotinylated bovine serum albumin immobilized onto each one of the bands. Due to the small inner volume of the capillary and the multianalyte feature we expect that the proposed device can be used for fast and inexpensive assays.