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

三名研究人员（阿肯色州大学医学科学的Storrie，约翰霍普金斯大学和本古里安大学的Brooker和罗森）在过去的7年里一直在合作，将全息术应用于荧光显微镜可以简化3D显微镜，并利用全息术提供的超分辨率功能。这项工作得到了NSF、NIH SBIR和NIST的资助赠款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。它是基于一个新的全息概念称为FINCH，开发的两个调查人员（罗森和布鲁克）。FINCH是一种全息技术，其中非相干荧光显微镜发射成像与一个简单的设置组成的空间光调制器插入到一个标准的荧光显微镜的立场，一个附加的数码相机的光路，和一个非激光，外部快门光源。最近发现FINCH技术具有固有的超分辨特性。该系统已发展到台式硬件和软件原型的阶段。在这项提案中，包括弗拉基米尔卢帕辛在内的三所大学的4名研究人员组成的网络， 阿肯色州医科大学将参与FINCH超分辨率显微镜技术的实施、研究使用和进一步改进三个方面。 1.在约翰霍普金斯大学（JHU），布鲁克和罗森将实现一个独立的FINCH显微镜，支持算法和图像采集和全息图重建软件将安装在UAMS在补助金年中期2。 2.在阿肯色州医学科学大学（UAMS），Storrie和Lupashin将利用FINCH显微镜推进膜运输和高尔基体组织的创新研究，否则这是不可能的。在这项工作中，他们还将比较FINCH显微镜的性能与其他超分辨率技术。 3.在本古里安大学（BGU），罗森将继续开发基于FINCH的先进算法和概念，以提高性能和分辨率。

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.