Fluorescence-based kinetic analysis of miniaturized protein microarrays

• Published in: Biosensors & bioelectronics 2018-12, Vol.122, p.290-299
• Main Authors: Ramiya Ramesh Babu, Heman Kumar, Gheber, Levi A.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Immobilized - chemistry; Biosensing Techniques - instrumentation; Chickens; Equipment Design; Fluorescence; Fluorescence detection; Immunoglobulin G - analysis; Kinetic parameters; Kinetics; Mice; Microfluidic; Microfluidic Analytical Techniques - instrumentation; Miniaturization; Multiplexed detection; Protein Array Analysis - instrumentation; Protein microarray; Rabbits
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2018.09.051

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. •Print miniaturized protein microarray, directly into a microchannel.•Image microarray in real-time, separate specific signal from noise.•Obtain real-time kinetic information from fluorescence-based detection.•Based on real time information, continuously estimate end-time of experiment.•Apply moving time-averaging to reduce background and increase SNR.