COBRE: UVM MED: CORE B: IMAGING & PHYSIOLOGY CORE

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。该设施主要用于活细胞成像和电生理学。它包含一个翻新的Noran/ Prairie Technologies激光扫描共聚焦显微镜，一个单级尼康TIRF（全内反射荧光）系统，一个比例成像系统，一个DeltaVision恢复显微镜系统和一个BioRad专用多光子激光扫描显微镜。我们也有膜片钳电子可用于大多数显微镜系统。Noran主要用于整个组织和分离细胞的钙成像。DeltaVision系统有一个温度和湿度控制的培养箱，允许对培养细胞进行延时实验，以及对固定样品进行免疫荧光研究。BioRad Radiance 2100专用多光子包括一个相干变色龙脉冲飞秒红外激光器和一个固定级直立奥林巴斯显微镜。红外激发，加上非扫描探测器，使得在500微米深处成像成为可能，持续数小时。它目前被用于脑切片工作。比率成像系统用于fura-2负载细胞或转染了基于fret的钙指示剂cameleon的细胞的成像。TIRF系统是一个相对较新的获得，但它将成为水泡运输研究和细胞粘附和迁移实验的工具。它也可用于近细胞膜的钙显像。对于数据反卷积和分析，我们有两个专用工作站运行softWoRx (Applied Precision), MetaMorph （Universal Imaging）和Volocity （Improvision）以及ImageJ （NIH）。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The facility is designed primarily for live-cell imaging and electrophysiology. It contains a refurbished Noran/ Prairie Technologies laser scanning confocal microscope, a single-level Nikon TIRF (total-internal reflection fluorescence) system, a ratiometric imaging system, a DeltaVision restoration microscopy system and a BioRad dedicated multiphoton laser scanning microscope. We also have patch-clamp electronics available for use on most microscope systems. The Noran is used primarily for calcium imaging on whole tissues and isolated cells. The DeltaVision system has a temperature and humidity-controlled incubator allowing for time-lapse experiments on cultured cells, as well as immunofluorescence studies on fixed samples. The BioRad Radiance 2100 Dedicated Multiphoton includes a Coherent Chameleon pulsed femtosecond IR laser and a fixed-stage upright Olympus microscope. The infrared excitation, coupled with non-descanned detectors, make it possible to image at depths of up to 500 microns for several hours. It is currently being used for brain-slice work. The ratiometric imaging system is used for imaging of fura-2 loaded cells or cells transfected with the FRET-based calcium indicator cameleon. The TIRF system is a relatively new acquisition, but it will become instrumental in vesicular trafficking studies and also cell adhesion and migration experiments. It also may be used for calcium imaging near the cell membrane. For data deconvolution and analysis, we have two dedicated workstations running softWoRx (Applied Precision) for deconvolution, MetaMorph (Universal Imaging), and Volocity (Improvision) along with ImageJ (NIH).