Hellen Dominick J, Karpen Saul J
Division of Pediatric Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Children's Healthcare of Atlanta and Emory University School of Medicine, Atlanta, GA, USA.
Bio Protoc. 2023 Jul 20;13(14):e4776. doi: 10.21769/BioProtoc.4776.
Current means to quantify cells, gene expression, and fibrosis of liver histological slides are not standardized in the research community and typically rely upon data acquired from a selection of random regions identified in each slide. As such, analyses are subject to selection bias as well as limited subsets of available data elements throughout the slide. A whole-slide analysis of cells and fibrosis would provide for a more accurate and complete quantitative analysis, along with minimization of intra- and inter-experimental variables. Herein, we present , a method for quantifying whole-slide scans of digitized histologic images to render a more comprehensive analysis of presented data elements. After loading images and preparing the project in the QuPath program, researchers are provided with one to two scripts per analysis that generate an average intensity threshold for their staining, automated tissue annotation, and downstream detection of their anticipated cellular matrices. When compared with two standard methodologies for histological quantification, had two significant advantages: increased speed and a 50-fold greater tissue area coverage. Using publicly available open-source code (GitHub), improves the reliability and reproducibility of experimental results while reducing the time scientists require to perform bulk analysis of liver histology. This analytical process is readily adaptable by most laboratories, requires minimal optimization, and its principles and code can be optimized for use in other organs. Graphical overview.
目前,在研究领域中,对肝脏组织学切片中的细胞、基因表达和纤维化进行量化的方法尚未标准化,通常依赖于从每张切片中选定的随机区域获取的数据。因此,分析容易受到选择偏差的影响,并且整个切片中可用数据元素的子集有限。对细胞和纤维化进行全切片分析将提供更准确和完整的定量分析,同时最大限度地减少实验内和实验间的变量。在此,我们提出了一种对数字化组织学图像的全切片扫描进行量化的方法,以便对所呈现的数据元素进行更全面的分析。在QuPath程序中加载图像并准备好项目后,研究人员每次分析会得到一到两个脚本,这些脚本会为他们的染色生成平均强度阈值、自动组织注释以及对预期细胞基质的下游检测。与两种组织学量化的标准方法相比,该方法有两个显著优点:速度提高,组织面积覆盖范围扩大50倍。使用公开可用的开源代码(GitHub),该方法提高了实验结果的可靠性和可重复性,同时减少了科学家对肝脏组织学进行大量分析所需的时间。这个分析过程大多数实验室都很容易采用,所需优化极少,其原理和代码可针对其他器官进行优化。图形概述。