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使用福尔马林/乙酸固定剂改进小鼠肠道组织的保存及使用QuPath进行定量组织学分析

Improved Preservation of Mouse Intestinal Tissue Using a Formalin/Acetic Acid Fixative and Quantitative Histological Analysis Using QuPath.

作者信息

Kandelouei Tahmineh, Zhang Wei, Houghton Madeline, Knudsen Beatrice, Edgar Bruce A

机构信息

Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah.

Department of Pathology, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah.

出版信息

Curr Protoc. 2024 May;4(5):e1062. doi: 10.1002/cpz1.1062.

DOI:10.1002/cpz1.1062
PMID:38775005
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11151780/
Abstract

The architecture and morphology of the intestinal tissue from mice or other small animals are difficult to preserve for histological and molecular analysis due to the fragile nature of this tissue. The intestinal mucosa consists of villi and crypts lined with epithelial cells. In between the epithelial folds extends the lamina propria, a loose connective tissue that contains blood and lymph vessels, fibroblasts, and immune cells. Underneath the mucosa are two layers of contractile smooth muscle and nerves. The tissue experiences significant changes during fixation, which can impair the reliability of histologic analysis. Poor-quality histologic sections are not suitable for quantitative image-based tissue analysis. This article offers a new fixative composed of neutral buffered formalin (NBF) and acetic acid, called FA. This fixative significantly improved the histology of mouse intestinal tissue compared to traditional NBF and enabled precise, reproducible histologic molecular analyses using QuPath software. Algorithmic training of QuPath allows for automated segmentation of intestinal compartments, which can be further interrogated for cellular composition and disease-related changes. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Improved preservation of mouse intestinal tissue using a formalin/acetic acid fixative Support Protocol: Quantitative tissue analysis using QuPath.

摘要

由于小鼠或其他小型动物的肠道组织性质脆弱，其结构和形态难以保存用于组织学和分子分析。肠黏膜由绒毛和隐窝组成，内衬上皮细胞。在上皮褶皱之间延伸的是固有层，这是一种疏松结缔组织，包含血管、淋巴管、成纤维细胞和免疫细胞。黏膜下方是两层收缩性平滑肌和神经。该组织在固定过程中会经历显著变化，这可能会损害组织学分析的可靠性。质量不佳的组织学切片不适用于基于图像的定量组织分析。本文提供了一种由中性缓冲甲醛（NBF）和乙酸组成的新型固定剂，称为FA。与传统的NBF相比，这种固定剂显著改善了小鼠肠道组织的组织学，并能够使用QuPath软件进行精确、可重复的组织学分子分析。QuPath的算法训练允许对肠道区域进行自动分割，进而可以进一步研究细胞组成和疾病相关变化。© 2024作者。由Wiley Periodicals LLC出版的《当前实验方案》。基本方案：使用甲醛/乙酸固定剂改善小鼠肠道组织的保存 支持方案：使用QuPath进行定量组织分析

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a01/11151780/784985e56b14/nihms-1992500-f0007.jpg
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