Biochip fluorescence detection system with spatial and spectral separation

Abstract

A novel optical arrangement for fluorescence detection that employs spatial separation as well as spectral filter to increase the signal to noise ratio is proposed. Using a prism and two mirrors, the elliptical laser beam of a laser diode, as an excitation light, is homogenized and the transmitted excitation light is separated from the fluorescence not to reach the collecting optics. Uncooled CCD can capture the fluorescence image of up to 40 fluorescently-labeled protein patterns without scanning or mechanical translation. This paper presents the simulation, construction and measurement results of the developed optical system. The measurements show that the combination of prism and mirrors converts the excitation light from the laser diode to uniform illumination on the specimen, and provides the separation between excitation and fluorescence light to give high signal to noise ratio. It is also possible to assay various protein concentrations ranging from 1000 to 10 ng/ml reliably. We believe that the proposed fluorescence detection system can be used to build a commercially valuable, low cost, hand-held or miniature fast detection device for point-of-care applications.