Fluorescence-based kinetic analysis of miniaturized protein microarrays

Ideal monitoring devices should enjoy a combination of characteristics, e.g. high sensitivity, multiplexing, portability, short time-to-result (TTR). Typically, no device meets all of these demands since some of them are contradictory, to some extent. Herein, we present a miniaturized platform based on fluorescent detection, which is sensitive, readily allows multiplexing, and allows real-time monitoring of the signal, thus allowing extraction of kinetic information as well as drastic reduction of TTR. This is achieved via miniaturization of active spots, integration with microfluidics, and algorithmic approaches. We validate its performance by comparing with evanescent field excitation, which obtains similar results, however without the addition of the necessary complex hardware.