Custom-built Digital Scanned Laser Light Sheet Microscope (DSLM)

定制数字扫描激光光片显微镜 (DSLM)

• 批准号: 8247214
• 负责人: KEVIN P. WHITE
• 金额: $ 50.93万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247214
• 关键词: Animal Model; Chicago; Complement; Coupled; Custom; Development; Disease Progression; Equipment; European; Expression Library; Fluorescence Microscopy; Funding; Genetic Transcription; Genomics; Grant; Housing; Image; Institutes; Laboratories; Lasers; Life; Light; Lighting; Microfluidic Microchips; Microscope; Microscopy; Minor; Molecular Biology; Noise; Organism; Physiological; Proteins; Rana; Research Infrastructure; Research Personnel; Resolution; Sampling; Scanning; Science; Signal Transduction; Specimen; Speed; System; Systems Biology; Techniques; Technology; Time; Tissues; United States National Institutes of Health; Universities; Zebrafish; digital; fly; innovation; instrument; programs; spatiotemporal; tissue/cell culture; two-photon

DESCRIPTION (provided by applicant): We request a custom built Digital Scanned Laser Light Sheet Microscope (DSLM), which will enable a broad group of investigators in Chicago to visualize proteins in live tissues and organisms with high resolution and speed and without sample damage associated with other fluorescence microscopy techniques. A major user group will be the investigators participating in at the Chicago Center for Systems Biology (CCSB) (://www.chicago-center-for-systems- biology.org/; grant P50GM081892). The Center's scientific program focuses on the dynamics of transcriptional networks on physiological, developmental and evolutionary scales. The new system will particularly enable us to generate and analyze extensive libraries of the expression networks in development, norm and disease progression. Because of the unique capabilities of this instrument, we have also received requests from groups outside The University of Chicago and we will reserve 25% time on the instrument for these users (minor user group). The DSLM is an innovative live-imaging fluorescence microscopy system developed by Dr. Stelzer's group at the European Molecular Biology Laboratory (EMBL) (Keller et al., 2008. Science, 322:1065-9). In comparison to other advanced fluorescence microscopy techniques (confocal and two-photon) the DSLM provides more than 50 times higher imaging speeds with 10 times higher signal to noise ratio, while exposing the specimens to at least two orders of magnitude less light. It presents a further improvement of a related technology, also introduced by the Stelzer group, known as Selective Plane Illumination Microscopy (SPIM) (Huisken et al., 2004. Science, 305:1007-9). The new equipment will be housed at the Institute for Genomics and Systems Biology (IGSB) (://www.igsb.org/) and will have an immediate impact on CCSB's advanced imaging platform for three important reasons. First, it would largely complement our existing confocal system coupled with a microfluidics device developed at The University of Chicago. Second, it will allow us for the first time to dynamically image fluorescently tagged transcription and other factors in model organisms (worm, fly, frog, and zebrafish) as well as in tissues and cell cultures under different physiological conditions over long periods of time, with the highest spatiotemporal resolution. Finally, the DSLM would leverage not only the CCSB research and infrastructure, but also other multiple NIH funded projects, including those presented in this application, bringing live imaging microscopy studies to the next level of speed and resolution. PUBLIC HEALTH RELEVANCE: The newly developed fluorescence Digital Scanned Laser Light Sheet Microscope (DSLM) dramatically minimizes photo damage to the specimen at the same time increasing both the speed and quality of live imaging. Such combination of features is unique to this instrument, which will enable investigators at The University of Chicago and collaborating institutions to perform in-vivo imaging experiments over long durations (e.g. several days of development)and at cellular resolution. This will greatly enhance our capabilities in cellular and developmental genomic research, particularly through visualizing and studying spatiotemporal networks of gene expression in norm and disease.

描述（由申请人提供）：我们要求定制的数字扫描激光光片显微镜（DSLM），这将使芝加哥的广泛研究人员能够以高分辨率和速度可视化活组织和生物体中的蛋白质，并且不会与其他荧光显微镜技术相关的样品损坏。一个主要的用户组将是参与芝加哥系统生物学中心（CCSB）的研究者（：//www.chicago-center-for-systems- biology.org/; grant P50 GM 081892）。该中心的科学计划侧重于生理，发育和进化尺度上的转录网络的动态。新系统将特别使我们能够生成和分析发育，规范和疾病进展中的表达网络的广泛库。由于该仪器的独特功能，我们还收到了来自芝加哥大学以外的团体的请求，我们将为这些用户（次要用户组）保留25%的仪器使用时间。DSLM是由欧洲分子生物学实验室（EMBL）的Stelzer博士的小组开发的创新的实时成像荧光显微镜系统（Keller等人，2008. Science，322：1065-9）。与其他先进的荧光显微镜技术（共聚焦和双光子）相比，DSLM提供了超过50倍的成像速度和10倍的信噪比，同时将样品暴露于至少两个数量级的光。它提出了相关技术的进一步改进，也是由Stelzer小组引入的，称为选择性平面照明显微术（SPIM）（Huisken等人，2004. Science，305：1007-9）。新设备将安置在基因组学和系统生物学研究所（IGSB）（：//www.igsb.org/），并将对CCSB的先进成像平台产生直接影响，原因有三个。首先，它将在很大程度上补充我们现有的共聚焦系统与芝加哥大学开发的微流体设备。其次，它将使我们第一次能够动态成像荧光标记的转录和其他因素在模型生物（蠕虫，苍蝇，青蛙和斑马鱼），以及在组织和细胞培养物在不同的生理条件下，在很长一段时间内，具有最高的时空分辨率。最后，DSLM不仅将利用CCSB的研究和基础设施，还将利用其他多个NIH资助的项目，包括本申请中提出的项目，将实时成像显微镜研究的速度和分辨率提高到一个新的水平。 公共卫生关系：新开发的荧光数字扫描激光光片显微镜（DSLM）大大减少了对样品的光损伤，同时提高了实时成像的速度和质量。这种功能组合是该仪器所独有的，这将使芝加哥大学的研究人员和合作机构能够在长时间内（例如，几天的开发）和细胞分辨率下进行体内成像实验。这将大大提高我们在细胞和发育基因组研究方面的能力，特别是通过可视化和研究正常和疾病中基因表达的时空网络。