Molecular and Cellular Imaging Core

分子和细胞成像和组织学核心（核心C）将为每个人提供三项主要服务， 本研究的三个项目是：1）成年小鼠心室肌细胞的分离和功能分析 (VC)，2）VC和心肌组织切片的显微镜分析，和3）VC和心肌组织切片中细胞死亡的评价。 心肌样本 1)VC的酶分离和电场刺激培养将在核心C中进行，以提供 可靠和可重复的风险投资，用于分析每个项目特定条件下的风险投资功能。 功能分析包括体外收缩性和细胞内钙（Ca 2+）瞬变的评估 分别使用数字化视频边缘检测系统和FURA 2检测。 2)许多基于显微镜的测定将可用于提供详细的免疫组织学和 荧光评估VC和心肌组织切片具体到每个项目。这些分析 包括VC和心肌组织切片、荧光 VC和心肌组织切片的分析和荧光/福斯特共振能量转移（FRET） 第二代信使和蛋白质相互作用的分析。 3)评价VC和心肌样本中的细胞死亡。核心C使用测定来检测细胞凋亡 和坏死细胞死亡。每个项目都将获得末端脱氧核苷酸转移酶的分析结果， 介导的dUTP缺口末端（TUNEL）标记、半胱天冬酶-3活性和膜联蛋白V染色分离的VC和/或 用于评价细胞凋亡的心肌切片/提取物，以及乙锭同型二聚体III（坏死）和 Hoechst（活细胞）VC染色和肌球蛋白重链染色作为指示剂或坏死， 心肌组织切片 所有四个项目将继续利用这一核心， 分离的VC，以及VC和心肌组织的高质量显微镜和细胞死亡分析。

The molecular and cellular imaging and histology core (Core C) will provide three main services to each of the three projects described: 1) isolation and functional analysis of murine adult ventricular cardiomyocytes (VC), 2) microscopy analysis of VC and myocardial tissue sections, and 3) evaluation of cell death in VC and myocardial samples. 1) Enzymatic isolation and electrical-field stimulated culture of VCs will be performed in Core C to provide reliable and reproducibly viable VC for analysis of VC function under conditions specific to each project. Functional analyses include assessment of in vitro contractility and intracellular calcium (Ca2+) transients using a digitalized video-edge detection system and FURA2 detection, respectively. 2) A number of microscopy-based assays will be available to provide detailed immunohistological and fluorescence assessment of VC and myocardial tissue sections specific to each project. These analyses include immunohistological and morphological analyses of VC and myocardial tissue sections, fluorescence analysis of VCs and myocardial tissue sections and fluorescence/forster resonance energy transfer (FRET) analysis of 2nd messenger generation and protein interactions in VCs. 3) To evaluate cellular death in VC and myocardial samples. Core C uses assays to detect both apoptotic and necrotic cell death. Each project will have access to analyses of terminal deoxynucleotidyl-transferase mediated dUTP nick end (TUNEL) labeling, caspase-3 activity and Annexin V staining isolated VC and/or myocardial sections/extracts for evaluation of apoptosis, as well as ethidium homodimer III (necrosis) and Hoechst (living cells) staining of VC and myosin heavy chain staining as an indicator or necrosis in myocardial tissue sections. This core will continue to be utilized by all four projects to provide reproducible functional analyses of isolated VC, as well as high quality microscopy and cell death analyses of VC and myocardial tissue.