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

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 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Nikon TIRF（全内反射荧光）系统，一个比率成像系统，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，可以对培养的细胞进行延时实验（如迁移试验），以及对固定样品进行免疫荧光研究。 BioRad Radiance 2100 Dedicated Multiphoton包括一个相干变色龙脉冲飞秒红外激光器和一个固定台直立Olympus显微镜。 红外激发，加上非去扫描探测器，使其能够在长达500微米的深度成像数小时。 它目前被用于脑切片中的钙成像和发育中鸡胚脊髓中神经元的逆行标记。 比率成像系统用于对负载fura-2的细胞或用基于FRET的钙指示剂cameleon转染的细胞进行成像。 TIRF系统用于囊泡运输研究以及细胞粘附和迁移实验。 将来，它也可能用于细胞膜附近的钙成像。 对于数据去卷积和分析，我们有两个专用的工作站，运行softWoRx（Applied Precision）进行去卷积，MetaMorph（Universal Imaging），Volocity（Improvement）和ImageJ（NIH）进行一般分析和渲染。

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 (such as migration assays), 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 calcium-imaging in brain-slices and retrograde labeling of neurons in the spinal cord of developing chick embryos. 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 used in vesicular trafficking studies and also cell adhesion and migration experiments. In the future, it may also 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), Volocity (Improvision) and ImageJ (NIH) for general analysis and rendering.