Imaging and Image Analysis Core

摘要 成像和图像分析核心将执行mRNA空间和细胞定位以及表达水平 分析，免疫荧光，以及定量标记的无纤维化成像对人类肺组织的基础上， 对每一个项目进行调查。来自广泛的特定功能性肺区域的样本 表型IPF肺代表了一种独特的资源，将使该计划中的所有三个项目受益， 标本需要仔细染色和成像，以进行mRNA和蛋白质的鉴定和定量 在一些实施方案中，细胞可以通过ER累积来评估ER应激、自噬、UPR、细胞衰老、细胞凋亡和纤维化。集中统一 先进的染色和显微成像技术也将为所有项目提供质量控制数据， 针对具体项目的调查，从而改善项目间协作和整合的机会。这 因此，核心是该方案的一个重要组成部分。 成像和图像分析核心的目标包括： ·设计染色组合并进行区域选择以进行全转录组分析， Nanostring GeoMX数字空间剖面仪。 ·对IPF和对照肺组织进行RNAscope原位杂交和显微镜成像， 补充，验证，并结合通过RNA-seq分析获得的转录谱， 以满足其他项目的具体需求。 ·对纤维状胶原进行无标记二次谐波生成成像以进行纤维化量化。 ·执行显微图像的分析和量化。 ·与数据管理和分析核心结合提供数字完整幻灯片图像存储 (Core D）。 成像和图像分析核心将由Evgenia Dobrinskikh博士领导，他将协调，概述， 为科学项目提供服务，并与其他研究中心协调。

ABSTRACT The Imaging and Image Analysis Core will perform mRNAs spatial and cellular localization and expression levels analyses, immunofluorescence, as well as quantitative label free fibrosis imaging on human lung tissues-based investigations for each one of the Projects. Samples from specific functional lung regions of extensively phenotyped IPF lungs represent a unique resource that will benefit all three Projects in the Program, and these specimens will need to be carefully stained and imaged for mRNA and protein identification and quantification accumulation to assess ER stress, autophagy, UPR, cell senescence, apoptosis, and fibrosis. Centralization of advanced staining and microscopic imaging will also benefit all Projects by providing quality-controlled data for Project-specific investigations, thus improving the opportunity for inter-project collaboration and integration. This Core is therefore an essential integrative component of the Program. The Aims of the Imaging and Image Analysis Core include: • Design staining combinations and perform regions selection for whole transcriptome analysis with Nanostring GeoMX Digital Spatial Profiler. • Perform RNAscope in situ hybridization and microscopic imaging of IPF and control lung tissue to complement, validate, and contextualize the transcriptional profiles obtained by RNA-seq analyses and to address other project-specific needs. • Perform label-free second harmonic generation imaging of fibrillar collagens for fibrosis quantification. • Perform analyses and quantification of microscopic images. • Provide Digital Whole Slide Images storage in conjunction with the Data Management and Analysis Core (Core D). The Imaging and Image Analysis Core will be led by Evgenia Dobrinskikh, PhD who will coordinate, overview, and perform services for the Scientific Projects and coordinate with other Research Cores.