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Device for laser microdissection

激光显微切割装置

基本信息

批准号：
447452855
负责人：
金额：
--
依托单位：
Universität des Saarlandes
依托单位国家：
德国
项目类别：
Major Research Instrumentation
财政年份：
2020
资助国家：
德国
起止时间：
2019-12-31 至无数据
项目状态：
未结题

来源：
https://gepris.dfg.de/gepris/projekt/447452855?language=en
关键词：
Device laser microdissection

项目摘要

The requested device for laser microdissection is to obtain high-purity samples from fixed and from living tissues without contamination in a semi-automatic manner. The samples including single cell samples will be characterized by miRNA, RNA, DNA sequencing (Institute of Human Genetics), by proteome and metabolome analysis (Institute of Pharmacology & Toxicology) and by further analytical assays including qPCR and Western Blotting (in all applicant groups). In contrast to the isolation of cells from dissociated tissues by cell sorting, the isolation of samples / cells from tissue sections with existing integrity of the cell network, allows the detection of paracrine signal mechanisms and accompanying changes of the transcriptome, the metabolome and the proteome. Specifically, laser microdissection will be used to analyze tissue-specific miRNA-mRNA interaction networks, in particular of tumor tissues, intracellular signaling pathways of endothelial cells in murine and human endometrial foci, and epithelial and mesenchymal single cells and immune cells from patient tissues and in vitro 3D cultures. Using gene-deficient tissues as controls, laser microdissection will also be used to investigated the effects of the absence of brush cells in the tracheal epithelium and the expression profile of defined tracheal epithelial cells in wild-type animals; the device will finally be used for targeting cells that contribute to the wound healing process of the skin (including immune cells, fibroblasts, macrophages, and vascular endothelia) and to non-infectiously triggered inflammatory processes of the liver (hepatocytes and pericytes).

所要求的激光显微解剖装置是以半自动的方式从固定组织和活体组织中获得无污染的高纯度样品。包括单细胞样本在内的样品将通过miRNA， RNA， DNA测序（人类遗传学研究所），蛋白质组和代谢组分析（药理学和毒理学研究所）以及包括qPCR和Western Blotting（所有申请人组）在内的进一步分析分析进行表征。与通过细胞分选从游离组织中分离细胞相比，从具有现有细胞网络完整性的组织切片中分离样品/细胞，可以检测旁分泌信号机制以及伴随的转录组、代谢组和蛋白质组的变化。具体来说，激光显微解剖将用于分析组织特异性miRNA-mRNA相互作用网络，特别是肿瘤组织，小鼠和人类子宫内膜病灶内皮细胞的细胞内信号通路，以及来自患者组织和体外3D培养的上皮和间充质单细胞和免疫细胞。以基因缺陷组织为对照，激光显微解剖还将用于研究野生型动物气管上皮中刷状细胞缺失的影响和明确的气管上皮细胞的表达谱；该装置最终将用于靶向有助于皮肤伤口愈合过程的细胞（包括免疫细胞、成纤维细胞、巨噬细胞和血管内皮细胞）和非感染性触发的肝脏炎症过程（肝细胞和周细胞）。