A Zeiss Elyra S1 Structured Illumination Microscope for the Biological Imaging Fa

描述（由申请人提供）：本提案的目的是为CNR生物成像设施（BIF）的卡尔蔡司Elyra S1 SIM系统获得NIH资助。这个超分辨率显微镜系统将增强BIF的研究和培训能力，特别是，将允许本提案中列出的六(6)个主要用户的研究目标继续进行他们积极的nih资助的研究。对于加州大学伯克利分校（379名pi /2431名用户）的许多科学家来说，BIF是涉及光学显微镜研究的主要焦点。然而，一个主要的限制是无法在“超分辨率”水平上解决荧光目标。目前，这些用户可用的最高分辨率光学成像技术是反褶积（API DeltaVision Elite）。校园里只有一种超分辨率系统（PALM）可用。我们的研究人员需要2-3通道的3D超分辨率成像，因此只有结构照明显微镜（SIM）适合我们的需求。STED不能与长波长探头一起工作，因此不适合我们所有的用户。PALM/STORM几乎没有3D信息。在本提案中，我们已经表明，由于缺乏解析小于衍射极限分辨率200nm的结构的能力，我们的主要用户的研究受到了损害。所要求的仪器将填补光学成像技术急需的空白，通过使用任何颜色探针提供可重复的100nm分辨率。这种关键能力将促进加州大学伯克利分校自然资源学院、文理学院和整个科学界的科学家的研究。在过去的一年中，我们使用卡尔蔡司Elyra S1，尼康N-SIM和API OMX对生物样品进行了初步的SIM实验，并将结果与使用BIF的反卷积和LSM 710共聚焦显微镜获得的结果进行了比较。对于每个主要用户的研究项目，我们都提供了明确的证据，表明SIM成像在分辨率，灵敏度和3D成像能力方面远远超过了我们现有显微镜的能力。

DESCRIPTION (provided by applicant): The purpose of this proposal is to acquire NIH funding for a Carl Zeiss Elyra S1 SIM system for the CNR Biological Imaging Facility (BIF). This super-resolution microscope system will augment the research and training capabilities of the BIF, and specifically, will permit the research goals of the six (6) Major Users listed in this proposal to proceed forward with their active NIH-funded research. The BIF is the main focus for research involving optical microscopy for a great many scientists at UC Berkeley (379 PIs/2431 users). A major limitation, however, is the inability to resolve fluorescent targets at a "super-resolution" level. At the present time the highest resolution optical imaging technique available to these users is deconvolution (API DeltaVision Elite). There is only one super-resolution system (PALM) available on campus. Our researchers require 2-3 channel 3D super-resolution imaging, and thus only a Structured Illumination Microscope (SIM) is appropriate for our needs. STED does not work with long wavelength probes and is therefore disqualified for all of our users. PALM/STORM yields little or no 3D information. In this proposal we have shown that the research of our Major Users is crippled by the lack of the ability to resolve structures smaller than diffraction-limited resolution of 200nm. The requested instrument will fill a much-needed gap in optical imaging technology by providing reproducible 100nm resolution using any color probe. This critical capability will facilitate the research of scientists in the College of Natura Resources, the College of Letters and Sciences, and the general scientific community at the University of California, Berkeley. Over the last year we performed a preliminary SIM experiments on biological samples using a Carl Zeiss Elyra S1, Nikon N-SIM, and the API OMX, and we compared the results to that obtained using the BIF's deconvolution and LSM 710 confocal microscopes. For each Major User's research project we present unequivocal evidence showing that SIM imaging far surpasses the capabilities of our existing microscopes in resolution, sensitivity, and the ability to image in 3D.