Planar FRET detection from biomolecules on an optofluidic chip

Autor: Brian S. Phillips, Aiqing Chen, Holger Schmidt, Philip Measor, Evan J. Lunt, Aaron R. Hawkins
Rok vydání: 2010
Předmět:
Zdroj: SPIE Proceedings.
ISSN: 0277-786X
DOI: 10.1117/12.841488
Popis: Fluorescence resonance energy transfer (FRET) is one of the powerful tools used to study the dynamics of biomolecules. By monitoring the energy transferred from a donor fluorophore to an acceptor fluorophore, one can determine the spatial proximity between the fluorophores on the nanometer scale and thus extract information regarding interactions between biomolecules. Here we demonstrate a novel way of measuring FRET from oligonucleotides using an integrated optofluidic chip containing a planar liquid-core waveguide that can guide liquid and light simultaneously. FRET experiments were carried out using fluorescein and Cy3 labeled oligonucleotides FRET pairs. An excitation laser was fiber-coupled to a solid-core waveguide perpendicular to the chip's liquid-core channel. A FRET efficiency of 50% was measured in good agreement with bulk microscopy experiments. By photobleaching the acceptors and manipulating the fluidic flow, we also demonstrated controllable FRET events: an increase in donor signal, a decrease in acceptor signal and the recovery of FRET due to the influx of new FRET pairs. The flexibility of our chip design also allows for improvements such as separate donor and acceptor detection at either chip end using integrated filters.
Databáze: OpenAIRE