Imaging and Image Analysis Core

成像和图像分析核心

• 批准号: 10627596
• 负责人: Evgenia Dobrinskikh
• 金额: $ 22.55万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10627596
• 关键词: Address; Apoptosis; Autophagocytosis; Biological Assay; Cell Aging; Collaborations; Collagen; Complement; Data; Data Analyses; Doctor of Philosophy; Fibrillar Collagen; Fibrosis; Fluorescence; Gene Expression Profile; Goals; Human; Image; Image Analysis; Immunofluorescence Immunologic; Immunohistochemistry; In Situ Hybridization; Individual; Investigation; Label; Lung; Messenger RNA; Mus; Pattern; Phenotype; Proteins; Pulmonary Fibrosis; Research; Resources; Sampling; Services; Specimen; Stains; Structure of parenchyma of lung; data management; design; digital; endoplasmic reticulum stress; improved; lung injury; microscopic imaging; nano-string; programs; second harmonic generation imaging; single-cell RNA sequencing; transcriptome; transcriptome sequencing; whole slide imaging

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.

抽象的 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 基于人体肺组织的分析、免疫荧光以及定量无标记纤维化成像 每个项目的调查样本主要来自特定的肺功能区域。 表型 IPF 肺是一种独特的资源，将使该计划中的所有三个项目受益，而这些 样本需要仔细染色和成像，以进行 mRNA 和蛋白质鉴定和定量 积累以评估 ER 应激、自噬、UPR、细胞衰老、细胞凋亡和纤维化。 先进的染色和显微成像也将通过提供质量控制数据使所有项目受益。 针对特定项目的调查，从而提高项目间协作和集成的机会。 因此，核心是该计划的重要组成部分。 成像和图像分析核心的目标包括： • 设计区域染色组合并使用以下方法进行全转录组分析的选择 Nanostring GeoMX 数字空间分析仪。 • 对 IPF 进行 RNAscope 原位杂交和显微成像并控制肺组织以 对通过 RNA-seq 分析获得的转录谱进行补充、验证和背景化 以满足其他项目特定的需求。 • 对纤维状胶原进行无标记二次谐波生成成像，以进行纤维化定量。 • 对显微图像进行分析和量化。 • 与数据管理和分析核心结合提供数字全幻灯片图像存储 （核心D）。 成像和图像分析核心将由 Evgenia Dobrinskikh 博士领导，她将协调、概述、 为科学项目提供服务并与其他研究核心进行协调。