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）：该共享仪器提案适用于配备专用于细菌延时成像的倒置落射荧光显微镜。用户是一群微生物学家，他们的研究重点是细菌生理学中的基本和突出问题。使用荧光显微镜对细菌进行细胞学分析在过去十年中已经成熟，并改变了细菌学领域。我们可视化亚细胞结构、细胞骨架元素甚至染色体定义区域的能力从根本上改变了我们对细菌细胞的看法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让我们思考这些分子如何发挥作用，还提供了检测方法来探索细胞分裂、染色体分离和形态发生等基本细胞生物过程的机制。此外，对含有荧光蛋白启动子融合的细胞的分析揭示了种群和群落中单个细菌细胞令人惊讶的变异和特化。使用荧光显微镜的许多革命性发现都来自于单个图像的采集（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，还有很多事情有待通过延时成像实时研究这些过程来发现。分析活细胞内蛋白质的动态行为；正在进行的形态发生过程；复杂群落中生长的细胞是细菌细胞生物学的下一个前沿领域。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、部门和整个校园中现有的大多数显微镜只能用于细菌细胞的基本延时成像。我们机构专门为延时显微镜设计的少数系统明显超额订购，并且缺乏对细菌大小的细胞进行成像所需的关键功能。拟议的专用于延时成像的荧光显微镜将加速当前的研究项目，实现发现并启动新项目。拟议的仪器将安置在哈佛医学院微生物学和分子遗传学系中心的一个开放式房间内。因此，它也将作为我们整个微生物学家社区的资源和培训场所。

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