Super-Res Holographic Microscopy to Advance Research on Golgi Apparatus Function

这一应用的三位研究者（阿肯色大学医学科学学院的Storrie，约翰霍普金斯大学和本古里安大学的Brooker和Rosen）在过去的7年里一直在合作，他们的愿景是将全息技术应用于荧光显微镜可以简化3D显微镜，并利用全息技术提供的超分辨率能力。这项工作得到了NSF、NIH SBIR和NIST的资助，现在已经产生了一种具有超分辨特性的三维荧光显微技术。它是基于一种叫做FINCH的新全息概念，由两位研究者（Rosen和Brooker）开发。FINCH是一种全息技术，通过一个简单的设置，包括插入到标准荧光显微镜支架的光路中的空间光调制器，附加的数码相机和非激光外部百叶窗光源，对非相干荧光显微镜发射进行成像。近年来，人们发现FINCH技术具有固有的超分辨特性。该系统已开发到一个台式硬件和软件样机阶段。在这项提议中，包括Vladimir Lupashin在内的三所大学的4名调查人员组成的网络

Three of the investigators on this application (Storrie at the University of Arkansas for Medical Sciences, Brooker and Rosen at Johns Hopkins University and Ben Gurion University) have been collaborating for the last 7 years on the vision that application of holography to fluorescence microscopy could simplify 3D microscopy and also capitalize on the super-resolution capabilities afforded by holography. This work, supported by NSF, NIH SBIR, and NIST grants hasnow yielded a 3-dimensional, fluorescence microscopic technique with super-resolving properties. It is based upon a new holography concept called FINCH, developed by two of the investigators (Rosen and Brooker). FINCH is a holographic technique in which the incoherent fluorescence microscope emission is imaged with a simple setup consisting of a spatial light modulator inserted into the light path of a standard fluorescence microscope stand, an attached digital camera, and a non-laser, externally shuttered light source. Recently the FINCH technique was found to have inherent super-resolving properties. This system has been developed to the stage of a bench-top hardware and software prototype. In this proposal, the network of 4 investigators at three universities including Vladimir Lupashin at University of Arkansas for Medical Sciences will participate in three aspects of the implementation, research use and further improvement ofthe FINCH super-resolution microscope technology. 1. At Johns Hopkins University (JHU), Brooker and Rosen will implement a stand-alone FINCH microscope, supporting algorithms and image acquisition and hologram reconstruction software to be installed at UAMS during the middle of grant Year 2. 2. At University of Arkansas for Medical Sciences (UAMS), Storrie and Lupashin will utilize the FINCH microscope to advance innovative research in membrane trafficking and Golgi organization that would not otherwise be possible. In that work, they will also compare FINCH microscope performance with other super-resolution techniques. 3. At Ben Gurion University (BGU), Rosen will continue to develop advanced algorithms and concepts based upon FINCH to improve performance and resolving power.