Biophotonics: Integrated Optical Coherence Tomography and Multi-Photon Microscopy for Microfluidic System Analysis and Molecular Beacon Monitoring

University Of Illinois At Urbana-Champaign, Urbana IL

Investigators

Abstract

0086696 Boppart The applicants propose to develop an integrated microscope capable of optical coherence tomography (OCT) and multi-photon microscopy (MPM) for simultaneous acquisition of microstructural and functional image data in microfluidic systems. This microscope will be used to investigate the design and performance of microfluidic mixing systems and to non-invasively monitor molecular beacon hybridization in real-time for the detection of specific nucleic acids from microbial pathogens and microorganisms in the environment. The microscope will enable the complementary use of OCT to provide microstructural and flow information and MPM to visualize the spatial distribution of fluorescent molecular beacons with respect to the microstructure. Optical coherence tomography is an emerging high-speed high-resolution imaging technology which has largely been applied to medical and biological applications. OCT performs optical ranging with micron-scale resolution in a manner similar to ultrasound, except reflections of near-infrared light are detected rather than sound. Optical Doppler OCT, a technique analogous to laser Doppler velocimetry, can acquire image-based data of fluid flow profiles and make quantitative measurements of flow velocities. The capabilities are ideally suited for diagnostic monitoring of three-dimensional microfluidic systems, which typically have microstructural dimensions of 10-100 mm. Molecular beacons, oligonucleotides that fluoresce only upon hybridization to a nucleic acid target site, are promising optical probes for the detection of microbial pathogens and other microbial populations. The optical sectioning capability of MPM will be used to simultaneously monitor the three-dimensional spatial distribution of fluorescing molecular beacons within microfluidic systems.