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，以使用抗原的双重和三重标记对各种蛋白质和细胞进行定位。用户组的研究兴趣是不同的，从调查胃肠道内的细胞多样性到心脏生理学，但都有共同的兴趣在测量细胞之间的空间并置，细胞转分化过程中蛋白质表达的变化，以及特定细胞类型内蛋白质的定位。用户组的研究记录非常好。用户组有相当丰富的经验，并发表了广泛使用共聚焦显微镜。然而，由于我们目前的显微镜（Biorad MRC 600）的局限性，年龄和广泛使用，目前的调查过程中出现的许多形态学问题不能得到充分解决。这对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移位。这些问题都需要对细胞表面或细胞内的抗原位点进行双重和三重标记。蔡司LSM 510激光共聚焦显微镜是严格完成已获资助项目的特定目标的理想工具。蔡司提供多达4个反射光共焦通道配置和一个透射光（DIC）共焦通道配置，多重跟踪关键荧光染料组合，单个可调针孔，用于正确的三维测量，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。内华达州大学的研究在过去的几年里有了显著的增长。蔡司LSM 510的拟议用户群竞争激烈，代表了内华达州大学一些最成功的研究人员。该机构通过每年为服务合同提供支持来承认用户群体的质量。PI在共聚焦显微镜方面拥有丰富的经验，将主持内部咨询委员会并监督仪器的日常操作。该仪器将被安置在现有的NIH资助的成像核心实验室。

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