Faculty of Biological and Environmental Sciences, University of Helsinki, Helsinki 00790, Finland.
Institute of Biotechnology, University of Helsinki, Helsinki 00790, Finland.
Cell Rep Methods. 2021 Jul 30;1(5):100059. doi: 10.1016/j.crmeth.2021.100059. eCollection 2021 Sep 27.
The intestine is divided into functionally distinct regions along the anteroposterior (A/P) axis. How the regional identity influences the function of intestinal stem cells (ISCs) and their offspring remain largely unresolved. We introduce an imaging-based method, "Linear Analysis of Midgut" (LAM), which allows quantitative, regionally defined cellular phenotyping of the whole midgut. LAM transforms image-derived cellular data from three-dimensional midguts into a linearized representation, binning it into segments along the A/P axis. Through automated multivariate determination of regional borders, LAM allows mapping and comparison of cellular features and frequencies with subregional resolution. Through the use of LAM, we quantify the distributions of ISCs, enteroblasts, and enteroendocrine cells in a steady-state midgut, and reveal unprecedented regional heterogeneity in the ISC response to a model of colitis. Altogether, LAM is a powerful tool for organ-wide quantitative analysis of the regional heterogeneity of midgut cells.
肠道沿前后（A/P）轴分为功能不同的区域。区域特征如何影响肠干细胞（ISCs）及其后代的功能在很大程度上仍未得到解决。我们引入了一种基于成像的方法，“中肠线性分析”（LAM），它允许对整个中肠进行定量的、区域定义的细胞表型分析。LAM 将源自三维中肠的图像衍生细胞数据转化为线性化表示，将其沿 A/P 轴划分为段。通过自动进行区域边界的多元确定，LAM 允许以亚区域分辨率对细胞特征和频率进行映射和比较。通过使用 LAM，我们量化了稳态中肠中 ISC、肠母细胞和肠内分泌细胞的分布，并揭示了 ISC 对结肠炎模型反应的前所未有的区域异质性。总之，LAM 是一种用于中肠细胞的区域异质性的器官范围定量分析的强大工具。
