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Proteins in biological fluids (blood, urine, cerebrospinal fluid) are important biomarkers of various pathological conditions. Protein biomarkers detection and quantification have been proven to be an indispensable diagnostic tool in clinical practice. There is a growing tendency towards using portable diagnostic biosensor devices for point-of-care (POC) analysis based on microfluidic technology as an alternative to conventional laboratory protein assays. In contrast to universally accepted analytical methods involving protein labeling, label-free approaches often allow the development of biosensors with minimal requirements for sample preparation by omitting expensive labelling reagents. The aim of the present work is to review the variety of physical label-free techniques of protein detection and characterization which are suitable for application in micro-fluidic structures and analyze the technological and material aspects of label-free biosensors that implement these methods. The most widely used optical and impedance spectroscopy techniques: absorption, fluorescence, surface plasmon resonance, Raman scattering, and interferometry, as well as new trends in photonics are reviewed. The challenges of materials selection, surfaces tailoring in microfluidic structures, and enhancement of the sensitivity and miniaturization of biosensor systems are discussed. The review provides an overview for current advances and future trends in microfluidics integrated technologies for label-free protein biomarkers detection and discusses existing challenges and a way towards novel solutions.

Abstract. The current contribution reports about the fabrication technology for the development of novel microfluidic impedance spectroscopy sensors that are directly attachable on 3-D molded interconnect devices (3D-MID) that provides an opportunity to create reduced-scale sensor devices for 3-D applications. Advantages of the MID technology in particular for an automotive industry application were recently discussed (Moser and Krause, 2006). An ability to integrate electrical and fluidic parts into the 3D-MID platform brings a sensor device to a new level of the miniaturization. The demonstrated sensor is made of a flexible polymer material featuring a system of electrodes that are structured on and embedded in the SU-8 polymer. The sensor chips can be directly soldered on the MID due to the electroless plated contact pads. A flip chip process based on the opposite electrode design and the implementation of all fluidic and electrical connections at one side of the sensors can be used to assemble the sensor to a three-dimensional substrate. The developed microfluidic sensor demonstrated a predictable impedance spectrum behavior and a sufficient sensitivity to the concentration of ethanol in deionized water. To the best of our knowledge, there is no report regarding such sensor fabrication technology.

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