Columbia AFM System Equipment Grant

描述(申请人提供)：为响应PAR-09-028：共享仪器资助计划(S10)，本申请申请支持最先进的生物显微镜催化原子力显微镜(AFM)系统(Veeco Instruments，Plainview，NY)，集成了新的奥林巴斯IX81倒置显微镜，配备了FluoView FV1000共聚焦激光扫描系统(LCSM，奥林巴斯，中央山谷，宾夕法尼亚州)，将安装在哥伦比亚大学生物医学工程系的核心设施中。提出这一要求的动机源于这样一个事实，即哥伦比亚大学目前没有核心设施(或我们所知的纽约市)为一般生命科学应用提供具有共焦能力的AFM系统。原子力显微镜已经成为生物医学工程研究的主要仪器，提供了从单分子到组织的重要成像和材料测试工具。集成系统将由多个NIH支持的研究人员在至少85%的时间内使用，并将在可用性的基础上供其他研究人员和学术目的访问。所要求的生物显微镜催化剂系统用户友好，集成了显微镜和分析软件(MIRO)，以及开放式扫描头设计，允许同时传输光应用，轻松获取样品，并可与FV1000 LSCM系统集成。结合这些成像系统的显微镜配置可用于细胞和组织级别的生物医学研究，其中AFM成像的结构可以与通过LCSM识别的生物分子相互关联。类似地，使用该系统，AFM力测量可以允许样本机械硬度的映射与LSCM可见的底层荧光标记的矩阵结构相关联。最后，使用激光共聚焦显微镜测量细胞对原子力显微镜诱导的机械刺激的反应，为机械生物学的研究提供了独特的机会，机械生物学研究细胞如何感知和响应他们的物理环境。FluoView FV1000 SIM(同时)模块进一步增强了这一集成AFM系统的功能，可提供光激活或漂白以及同步图像采集，从而实现光漂白后荧光恢复(FRAP)和分子去除应用。因此，一个提供AFM和LSCM相结合能力的研究平台将为生物应用提供一个强大的研究工具。BME部门是市中心校区最多的NIH支持的调查人员的所在地，并已做好准备领导这项共享仪器的努力，进一步架起两个校区(市中心晨边高地到上城医学院)的桥梁，并促进哥伦比亚大学的跨学科研究合作。

DESCRIPTION (provided by applicant): In response to PAR-09-028: Shared Instrumentation Grant Program (S10), this application requests support for a state-of-the-art BioScope Catalyst Atomic Force Microscope (AFM) system (Veeco Instruments, Plainview, NY) integrated with a new Olympus IX81 inverted microscope equipped with a Fluoview FV1000 confocal laser scanning system (LCSM, Olympus, Center Valley, PA) to be housed in the core facilities of the Department of Biomedical Engineering at Columbia University. Motivation for this request stems from the fact that there are currently no core facilities at Columbia University (or that we are aware of in New York City) that provide an AFM system with confocal capabilities for general life sciences applications. The AFM has become a staple instrument for biomedical engineering research, providing an important imaging and material testing tool spanning applications from single molecules to tissues. The integrated system will be used by multiple NIH- supported investigators for at least 85% of the time and will be made accessible for other researchers and academic purposes on an availability basis. The requested BioScope Catalyst system is user-friendly with integrated microscopy and analyses software (MIRO), as well as an open scan head design that permits simultaneous transmitted light applications, easy sample access and can be integrated with the FV1000 LSCM system. A microscope configuration combining these imaging sytems can be used to great advantage in cellular and tissue-level biomedical research where structures imaged with AFM can be cross-correlated to biomolecules identifiable via the LCSM. Similarly, using this system, AFM force measurements can permit mapping of sample mechanical stiffness to be correlated with underlying fluorescently labeled matrix structures visible by LSCM. Lastly, measurement of cellular responses using LCSM in response to AFM-induced mechanical stimulation provides unique opportunities for studies of mechanobiology, which investigates how cells perceive and respond to their physical environment. Further enhancing the capabilities of this integrated AFM system, the Fluoview FV1000 SIM (SIMultaneous) module provides photoactivation or bleaching and simultaneous image collection, enabling fluorescence recovery after photobleaching (FRAP) and molecular uncaging applications. Therefore, a research platform that delivers combined AFM and LSCM capabilities would offer a powerful research tool for biological applications. The BME department is home to the largest number of NIH-supported investigators at the downtown campus and is well-poised to lead this Shared Instrumentation effort, serving to further bridge the two campuses (downtown Morningside Heights to the uptown Medical Campus) and foster interdisciplinary research collaborations at Columbia.