Authors: B. Filanoski, S. Rastogi, E. Cameron, N. Mishra and W. Maki
Affilation: University of Idaho, United States
Pages: 453 - 456
Keywords: bioluminescence, BRET, Imaging chip, microarray, bio-detection
Fluorescent microarray technologies are widely used in molecular biology for studying gene expression, immune detection and drug discovery. Fluorescent assays are very sensitive but present problems with many bio-agents having inherent fluorescence which interfere with assays. In addition, auto-fluorescence and photo-bleaching are common problems in fluorescent detection. Moreover, fluorescence detection requires an expensive external light source coupled to expensive detection equipment. In the field of clinical diagnostics a sensitive and rapid homogeneous immunoassay is needed for harmful bio-agent detection. This paper proposes a rapid and potentially sensitive micro-chip detector based on Bioluminescence Resonance Energy Transfer (BRET). Bio-luminescent assays are exquisitely sensitive and not limited by overlap between fluorescent properties of assay components, and achieve a high signal to noise ratio. To achieve high spatial resolution, a new homogeneous bioluminescence resonance energy transfer method for imaging chip based high density array technology is presented. The distance between donor and acceptor as a way to conjugate fluorescent dyes directly to alkaline phosphatase is minimized in the new work. As a result, the efficiency of resonant energy transfer and achieved spatial resolution is increased. This method allows direct measurement of a fluorescent signal on an imaging chip without conventional optical instruments.