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

申请人建议开发一种能够进行光学相干层析成像(OCT)和多光子显微镜(MPM)的集成显微镜，用于同时采集微流体系统中的微结构和功能图像数据。该显微镜将用于研究微流控混合系统的设计和性能，并实时非侵入性地监测分子信标杂交，以检测环境中微生物病原体和微生物的特定核酸。显微镜将补充使用OCT来提供微观结构和流动信息，以及MPM来可视化荧光分子信标相对于微结构的空间分布。光学相干层析成像是一种新兴的高速高分辨率成像技术，已广泛应用于医学和生物领域。OCT以类似于超声波的方式执行微米级分辨率的光学测距，只是检测到的是近红外光的反射，而不是声音。光学多普勒OCT是一种类似于激光多普勒测速的技术，它可以获取基于图像的流体流动剖面数据，并对流动速度进行定量测量。这些功能非常适合三维微流控系统的诊断监测，这些系统通常具有10-100 mm的微结构尺寸。分子信标是一种只有在杂交到核酸靶点时才会发出荧光的寡核苷酸，是检测微生物病原体和其他微生物种群的很有前途的光学探针。MPM的光学切片能力将被用来同时监测微流控系统中荧光分子信标的三维空间分布。

0086696BoppartThe 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.