Combined Optical Tweezers-Fluorescence Super-Resolution Microscope for Single-Molecule Biophysical Studies

项目总结 在这一应用中，我们提出了一种组合式光学镊子-荧光超分辨显微镜 用于LUMICKS的单分子生物物理研究。LUMICKS C-Trap将放置在共享的 仪器设施，使其可供整个生物医学研究社区使用，网址为 哥伦比亚大学。因此，C陷阱将影响目前和未来由NIH资助的大量 哥伦比亚大学的研究项目。具体地说，C型疏水阀将被安装、容纳、操作和 保存在哥伦比亚大学精密生物分子表征设备(PBCF)，一所大学- 支持、配备人员、共享的仪器设施，将为所有哥伦比亚大学的研究人员提供 高效、低成本地使用C-Trap。拟议的C-Trap是一种最先进的仪器，将是 配备四脉冲激光镊子，三波长，单光子敏感的激光扫描共焦 荧光显微镜和受激发射耗尽(STED)超分辨荧光显微镜 (即，STED纳米显微镜)，从而能够容纳最广泛的研究。确实有 目前在哥伦比亚大学没有类似的仪器，只有两个C-Trap 整个纽约市地区的乐器。这两个仪器不位于共享仪器中 设施，取而代之的是位于个人研究人员的实验室，在那里仪器只 服务于个人研究人员实验室的实验需求，因此不可用 提供给外部用户。此外，这两个仪器的配置与请求的仪器不同，并且 能力有限，因此不能用于执行当前建议的实验 申请。在PBCF中成功建立拟议的C-Trap有望推进更多目标 超过26个由美国国立卫生研究院资助的研究项目，涉及哥伦比亚大学和 代表了极其广泛的生化、生物物理和生物医学研究项目。

PROJECT SUMMARY In this application, we propose to acquire a combined optical tweezers-fluorescence super-resolution microscope for single-molecule biophysical studies from LUMICKS. The LUMICKS C-Trap will be placed in a shared instrumentation facility such that it will be available for use by the entire biomedical research community at Columbia University. Consequently, the C-Trap will impact a large number of current and future NIH-funded research programs at Columbia University. Specifically, the C-Trap will be installed, housed, operated, and maintained in the Columbia University Precision Biomolecular Characterization Facility (PBCF), a university- supported, staffed, shared instrumentation facility that will provide all Columbia University researchers with efficient, low-cost access to the C-Trap. The proposed C-Trap is a state-of-the-art instrument that will be equipped with quadrupule laser tweezers, a three-wavelength, single-photon-sensitivity laser scanning confocal fluorescence microscope, and stimulated emission depletion (STED) super-resolution fluorescence microscope (i.e., a STED nanoscope), thereby enabling the broadest range of research to be accommodated. There are currently no comparable instruments anywhere at Columbia University and there are only two C-Trap instruments in the entire New York City area. These two instruments are not located in a shared instrumentation facility and are instead located in the laboratory of individual researchers, where the instruments exclusively serve the experimental needs of the individual researcher’s laboratory and are therefore rendered unavailable to outside users. Moreover, these two instruments are configured differently from the requested instrument and have limited capabilities and, thus, cannot be used to perform the experiments proposed in the current application. Successfully establishing the proposed C-Trap in the PBCF is expected to advance the aims of more than 26 NIH-funded research programs spanning many departments and schools at Columbia University and representing an extremely broad range of biochemical, biophysical, and biomedical research programs.