MRI: Development of high-throughput light sheet fluorescence microscopy

MRI：高通量光片荧光显微镜的发展

• 批准号: 1427957
• 负责人: Raghuveer Parthasarathy
• 金额: $ 30.56万
• 依托单位: University of Oregon Eugene
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1427957&HistoricalAwards=false
• 关键词: MRI; Development; throughput; light; sheet

An award is made to the University of Oregon to develop an instrument capable of three-dimensional imaging of large numbers of living multicellular specimens, especially embryos and larvae of zebrafish that serve as models for the study of many biological processes. Observation of cellular interactions during early development, wound healing, disease progression, and many other events has been enormously beneficial to our understanding of both normal and aberrant biological function. We have lacked methods, however, to apply recently developed advanced three-dimensional imaging techniques to large numbers of specimens, limiting both the rate of experiments and the statistical robustness of results. This project addresses this challenge through the development of new instrumentation. Implementing this research will provide valuable cross-disciplinary training for graduate students and undergraduates, as it fosters the development of skills in optics, device control, computational data analysis, microbiology, and developmental biology. In addition to research, the instrument will be used in activities related to summer undergraduate research internships and a day camp that the PI has co-organized for several years that exposes socioeconomically disadvantaged high school students to science. More broadly, the instrumentation developed by this project will provide previously unseen views of cells, tissues, and organs in living animals that can underpin new perspectives on health and disease.In recent years, advanced three-dimensional imaging methods have provided deep insights into the spatial and temporal arrangements of cells. A high-resolution, three-dimensional microscopy method known as light sheet fluorescence microscopy has proven itself to be particularly powerful for imaging living specimens over many-hour or several-day durations. This and similar methods, however, have suffered the limitation of very limited throughput, capable of imaging one or at most a few specimens during an experimental run. It is increasingly apparent that the high variability of developing organisms, and the subtleties of mechanisms governing them, call for studies that can apply high-precision imaging to much larger numbers of specimens. This motivates the instrument we will design and build, which integrates high-throughput specimen handling and real-time computational image analysis with light sheet fluorescence microscopy. Specific imaging targets that we will design our instrument to be capable of imaging and analyzing include the gut microbiota, immune system, and groups of neurons. Beyond this, there is a large list of systems for which long-duration three-dimensional imaging with high statistical power would be useful, implying a sizeable impact for our instrumentation.

俄勒冈州大学获得一项奖励，以开发一种能够对大量活的多细胞标本进行三维成像的仪器，特别是作为许多生物过程研究模型的斑马鱼胚胎和幼体。在早期发育、伤口愈合、疾病进展和许多其他事件中观察细胞相互作用对我们理解正常和异常的生物学功能都是非常有益的。然而，我们缺乏方法，将最近开发的先进三维成像技术应用于大量标本，限制了实验的速度和结果的统计稳健性。该项目通过开发新的仪器来应对这一挑战。实施这项研究将为研究生和本科生提供有价值的跨学科培训，因为它促进了光学，设备控制，计算数据分析，微生物学和发育生物学技能的发展。 除了研究之外，该工具还将用于与夏季本科生研究实习和PI多年来共同组织的日间夏令营有关的活动，这些活动使社会经济弱势高中生接触科学。更广泛地说，该项目开发的仪器将提供以前从未见过的活体动物细胞，组织和器官的视图，可以支持健康和疾病的新视角。近年来，先进的三维成像方法已经为细胞的空间和时间排列提供了深刻的见解。被称为光片荧光显微镜的高分辨率三维显微镜方法已被证明是特别强大的成像活标本超过数小时或数天的持续时间。然而，这种方法和类似的方法受到非常有限的吞吐量的限制，能够在实验运行期间对一个或至多几个标本进行成像。越来越明显的是，发育中的生物体的高度变异性，以及控制它们的机制的微妙性，要求研究能够将高精度成像应用于更大数量的标本。这激发了我们将设计和建造的仪器，该仪器将高通量样品处理和实时计算图像分析与光片荧光显微镜相结合。我们将设计能够成像和分析的仪器的特定成像目标包括肠道微生物群，免疫系统和神经元组。 除此之外，还有大量的系统，其中具有高统计功效的长时间三维成像将是有用的，这意味着对我们的仪器有相当大的影响。