Multipoint parallel excitation and CCD-based imaging system for high-throughput fluorescence detection of biochip micro-arrays

Abstract

We report the development and the characterization of a multipoint parallel excitation and CCD-based imaging system for high-throughput fluorescence detection of biochip micro-arrays. A two-dimensional array of (19×19) points with uniform intensity distribution, generated by a holographic array generator, was used for parallel excitation of two-dimensional micro-arrays of fluorescence samples. A CCD-based imaging system was used for high-throughput parallel detection and quantitative analysis of the fluorescence output. Micro-array samples of cyanine (Cy5) dye dots on silicon wafers and on glass substrates with varying concentration were used to evaluate the performance of the system. Results of fluorescence intensity measurements with varying concentration of dye and with different image acquisition time are presented. We have demonstrated that this novel approach will, in general, outperform the conventional approach in the excitation efficiency, the signal-to-noise ratio, and the throughput. The limitations and the potential improvements of the present method are discussed.