Zeiss LSM 510 Laser Scanning Confocal Microscope

蔡司 LSM 510 激光扫描共焦显微镜

• 批准号: 6440771
• 负责人: Sean M Ward
• 金额: $ 45.25万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440771
• 关键词: biomedical equipment purchase; cell differentiation; confocal scanning microscopy; gene expression; protein localization

This grant proposal seeks funding for a laser scanning confocal microscope to perform localization of a variety of proteins and cells using double and triple labeling of antigens. The research interests of the user group are diverse ranging from investigating cellular diversity within the gastrointestinal tract to cardiac physiology, but all have common interests in measuring the spatial apposition between cells, changes in protein expression during cellular transdifferentiation, and the localization of proteins within specific cell types. The research record of the user group is excellent. The user group has considerable experienced and has published extensively using confocal microscopy. However, because of limitations, age and extensive use of our current microscope (Biorad MRC 600) many of the morphological questions that have arisen during the course of current investigations cannot be adequately addressed. This is a serious limitation for the ongoing projects of the user group. Questions that need to be addressed include an examination of the spatial relationships between cells in heterogeneous tissues, the co- localization of organelles and proteins and translocation of proteins within cells. These are questions that all require double and triple labeling of antigen sites on the surface or within cells. The ideal instrument to facilitate rigorous completion of specific aims of already funded projects to incorporate a Zeiss LSM 510 laser confocal microscope. The Zeiss provides up to 4 confocal channel configurations for reflected light plus one for transmitted light (DIC), multi-tracking for critical flurochrome combinations, individual adjustable pinholes for correct 3-dimensional measurements, 12 bit data acquisition and 2048x2048 pixel resolution. The Zeiss offers the greatest integrated system available for these studies. Research at the University of Nevada has grown dramatically over the last number of years. The proposed user group of the Zeiss LSM 510 is highly competitive, representing some of the most successful investigators at the University of Nevada. The institution recognizes the quality of the user group by providing yearly support for a service contract. The PI has had considerable experience with confocal microscopy and will chair the internal advisory committee and oversee the daily operation of the instrument. The instrument will be housed in an existing NIH funded imaging core laboratory.

该拨款提案寻求资助激光扫描共聚焦显微镜，以使用双标记和三标记抗原对各种蛋白质和细胞进行定位。用户组的研究兴趣多种多样，从研究胃肠道内的细胞多样性到心脏生理学，但所有人都有共同的兴趣，包括测量细胞之间的空间排列、细胞转分化过程中蛋白质表达的变化以及蛋白质在特定细胞类型内的定位。用户组的研究记录非常出色。用户组拥有丰富的经验，并使用共焦显微镜发表了大量文章。然而，由于我们当前的显微镜（Biorad MRC 600）的局限性、年龄和广泛使用，当前研究过程中出现的许多形态学问题无法得到充分解决。这对于用户组正在进行的项目来说是一个严重的限制。需要解决的问题包括检查异质组织中细胞之间的空间关系、细胞器和蛋白质的共定位以及细胞内蛋白质的易位。这些问题都需要对表面或细胞内的抗原位点进行双重和三重标记。理想的仪器，可促进严格完成已资助项目的特定目标，以纳入蔡司 LSM 510 激光共焦显微镜。 Zeiss 提供多达 4 个用于反射光的共焦通道配置和一个用于透射光 (DIC) 的共焦通道配置、用于关键荧光染料组合的多跟踪、用于正确 3 维测量的单独可调针孔、12 位数据采集和 2048x2048 像素分辨率。蔡司为这些研究提供了最好的集成系统。内华达大学的研究在过去几年中取得了巨大的发展。 Zeiss LSM 510 的拟议用户组竞争非常激烈，代表了内华达大学一些最成功的研究人员。该机构通过提供年度服务合同支持来认可用户群体的质量。 PI 在共焦显微镜方面拥有丰富的经验，并将担任内部咨询委员会的主席并监督仪器的日常运行。该仪器将安置在 NIH 资助的现有成像核心实验室中。

项目成果

Calcium transients in intramuscular interstitial cells of Cajal of the murine gastric fundus and their regulation by neuroeffector transmission.

小鼠胃底卡哈尔肌内间质细胞中的钙瞬变及其神经效应器传递的调节。

• DOI: 10.1113/jp282876
• 发表时间: 2022
• 期刊: The Journal of physiology
• 作者: Hwang,SungJin;Drumm,BernardT;Kim,MinKyung;Lyu,JuHyeong;Baker,Sal;Sanders,KentonM;Ward,SeanM
• 通讯作者: Ward,SeanM