Design, simulation and fabrication of a total internal reflection (TIR)-based chip for highly sensitive fluorescent imaging

Abstract

This paper presents a total internal reflection based chip which generates evanescent waves for highly sensitive fluorescent imaging. The chip is monolithically, massively cast in polydimethylsiloxane (PDMS) at a very low cost using a Si mold fabricated by Si anisotropic wet etching and deep reactive ion etching (DRIE). Our method integrates all miniaturized optical components, namely cylindrical microlens, prism and waveguide, into one monolithic PDMS chip; thus assembly is unnecessary, and misalignment is eliminated. The slide-format and monolithic chip can be used with both upright and inverted fluorescent microscopes with flexible sample delivery platforms. The flexibility of sample delivery platforms facilitates various surface treatment/immobilization techniques required in fluorescent imaging. Moreover, the fiberoptics coupling into the chip allows a broad choice of wavelengths and types of laser sources ranging from UV to IR. We have successfully demonstrated the capability of the chip in highly sensitive imaging of tetramethylrhodamine (TMR) fluorescent dye and immobilized fluorescent nanobeads. Our monolithic, miniaturized TIR-based chip could potentially serve as an evanescent excitation-based platform integrated into a micro-total analysis system (μ-TAS).