Fluorescence Microscope for Time-Lapse Imaging of Bacteria

用于细菌延时成像的荧光显微镜

• 批准号: 7792067
• 负责人: DAVID Z RUDNER
• 金额: $ 16.2万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-22 至 2011-10-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792067
• 关键词: Age; Bacteria; Bacterial Physiology; Bacteriology; Behavior; Biological Assay; Biological Process; Cell Size; Cell division; Cell physiology; Cells; Cellular biology; Chromosome Segregation; Chromosomes; Communities; Complex; Elements; Fluorescence Microscopy; Heart; Housing; Image; Individual; Institution; Laboratories; Life; Microbiology; Microscope; Microscopy; Molecular Genetics; Morphogenesis; Perception; Population; Process; Proteins; Research; Research Project Grants; Resources; Site; Subcellular structure; System; Thinking; Time; Training; Variant; cell type; design; fluorescence microscope; frontier; instrument; medical schools; promoter

DESCRIPTION (provided by applicant): This shared instrument proposal is for an inverted epifluorescence microscope outfitted for, and dedicated to, time-lapse imaging of bacteria. The users are a group of microbiologists whose research is focused on fundamental and outstanding problems in bacterial physiology. Cytological analysis of bacteria using fluorescence microscopy has come of age over the last decade and has transformed the field of bacteriology. Our ability to visualize subcellular structures, cytoskeletal elements, and even defined regions of the chromosome has fundamentally changed our perception of the bacterial cell and the types and depth of questions we ask about it. Subcellular localization of proteins not only informs our thinking about how these molecules function it also provides assays to probe the mechanisms underlying basic cell biological processes like cell division, chromosome segregation and morphogenesis. In addition, the analysis of cells harboring promoter fusions to fluorescent proteins has revealed surprising variation and specialization of individual bacterial cells within populations and communities. Many of the revolutionary discoveries using fluorescence microscopy have come from the acquisition of single images (called "snap-shot" microscopy). It is becoming increasingly clear that these snap shots are only telling part of the story and that much remains to be discovered by studying these processes in real time by time- lapse imaging. Analysis of the dynamic behavior of proteins inside living cells; of ongoing morphogenetic processes; and of cells growing within complex communities is the next frontier in bacterial cell biology. The proposed instrument will help us explore this frontier and stimulate the next wave of breakthroughs. Most of the existing microscopes in our laboratories, our department, and across our campus can only be used for rudimentary time-lapse imaging of bacterial cells. The few systems at our institution specifically designed for time-lapse microscopy are significantly over-subscribed and lack the critical features necessary for imaging cells the size of bacteria. The proposed fluorescent microscope dedicated to time-lapse imaging would accelerate current research projects, enable discovery, and launch new projects. The proposed instrument will be housed in an open-access room in the heart of the department of Microbiology and Molecular Genetics at Harvard Medical School. Accordingly, it will also serve as a resource and training site for our entire community of microbiologists.

描述（由申请人提供）：这个共享的仪器提案是一个倒置的荧光显微镜，专门用于细菌的延时成像。用户是一群微生物学家，他们的研究重点是细菌生理学的基础和突出问题。在过去的十年里，利用荧光显微镜对细菌进行细胞学分析已经成熟，并改变了细菌学领域。我们对亚细胞结构、细胞骨架元素、甚至染色体特定区域的可视化能力，已经从根本上改变了我们对细菌细胞的认知，以及我们提出的问题的类型和深度。蛋白质的亚细胞定位不仅告诉我们这些分子是如何起作用的，而且还提供了探测基本细胞生物学过程（如细胞分裂、染色体分离和形态发生）的机制的方法。此外，对携带启动子与荧光蛋白融合的细胞的分析揭示了群体和群落中单个细菌细胞的惊人变化和特化。使用荧光显微镜的许多革命性发现都来自单幅图像的获取（称为“快照”显微镜）。越来越清楚的是，这些快照只是讲述了故事的一部分，还有很多有待于通过延时成像实时研究这些过程来发现。活细胞内蛋白质动态行为分析；正在进行的形态发生过程的；细胞在复杂群落中生长是细菌细胞生物学的下一个前沿。拟议的工具将帮助我们探索这一前沿，并激发下一波突破。我们实验室里，我们系里，校园里现有的大多数显微镜只能用来对细菌细胞进行基本的延时成像。我们机构专门为延时显微镜设计的为数不多的系统明显超额订购，并且缺乏成像细菌大小的细胞所需的关键特征。拟议中的用于延时成像的荧光显微镜将加速当前的研究项目，使发现成为可能，并启动新的项目。拟议中的仪器将被安置在哈佛医学院微生物学和分子遗传学中心的一个开放的房间里。因此，它也将作为我们整个微生物学家社区的资源和培训网站。

项目成果

A central role for PBP2 in the activation of peptidoglycan polymerization by the bacterial cell elongation machinery.

• DOI: 10.1371/journal.pgen.1007726
• 发表时间: 2018-10
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Rohs PDA;Buss J;Sim SI;Squyres GR;Srisuknimit V;Smith M;Cho H;Sjodt M;Kruse AC;Garner EC;Walker S;Kahne DE;Bernhardt TG
• 通讯作者: Bernhardt TG