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灯板显微镜（DSLM），这将使芝加哥的大量研究人员能够可视化活组织和生物体中具有高分辨率和速度的生物体中的蛋白质，并且没有样品损害与其他荧光显微镜技术相关的样品损害。主要用户组将是参加芝加哥系统生物学中心（CCSB）（：//www.chicago-center-for-systems-systems-biology.org/; Grant P50GM081892）的研究人员。该中心的科学计划着重于针对生理，发育和进化量表的转录网络的动态。新系统将特别使我们能够在发展，规范和疾病进程中生成和分析表达网络的广泛图书馆。由于该乐器具有独特的功能，我们还收到了芝加哥大学以外的小组的要求，我们将为这些用户（次要用户组）保留25％的时间。 DSLM是由Stelzer博士组在欧洲分子生物学实验室（EMBL）开发的创新现场荧光显微镜系统（Keller等，2008。Science，322：1065-9）。与其他晚期荧光显微镜技术（共聚焦和两光子）相比，DSLM提供的成像速度高50倍以上，同时将标本暴露于至少两个幅度较小的光级。它提出了相关技术的进一步改进，该技术也由Stelzer组引入，称为选择性平面照明显微镜（SPIM）（Huisken等，2004。Science，305：1007-9）。新设备将安置在基因组和系统生物学研究所（IGSB）（：//www.igsb.org/），并出于三个重要原因将对CCSB的高级成像平台产生直接影响。首先，它将在很大程度上补充我们现有的共聚焦系统，再加上芝加哥大学开发的微流体设备。其次，这将使我们首次在模型生物（蠕虫，苍蝇，青蛙和斑马鱼）以及在不同生理条件下在不同的生理条件下，在模型生物（蠕虫，蝇，青蛙和斑马鱼）中动态图像荧光标记的转录和其他因素，并在不同的生理条件下，具有最高的时空分辨率。最后，DSLM不仅​​会利用CCSB的研究和基础设施，还利用其他多个NIH资助的项目，包括本应用程序中提出的项目，将实时成像显微镜研究提升到更高的速度和分辨率水平。 公共卫生相关性：新开发的荧光数字扫描激光光显微镜（DSLM）大大最大程度地减少了对样品的照片损害，同时又提高了实时成像的速度和质量。这种功能的组合是该工具独有的，它将使芝加哥大学的研究人员和合作机构能够在长时间（例如开发几天）和蜂窝分辨率上进行体内成像实验。这将大大增强我们在细胞和发育基因组研究中的能力，特别是通过可视化和研究规范和疾病中基因表达的时空网络。