Virtual unfolding of light sheet fluorescence microscopy dataset for quantitative analysis of the mouse intestine

被引:17
作者
Candeo, Alessia [1 ]
Sana, Ilenia [2 ]
Ferrari, Eleonora [2 ]
Maiuri, Luigi [2 ,3 ]
D'Andrea, Cosimo [1 ]
Valentini, Gianluca [1 ]
Bassi, Andrea [1 ]
机构
[1] Politecn Milan, Dipartimento Fis, Piazza Leonardo da Vinci 32, I-20133 Milan, Italy
[2] Osped San Raffaele, IERFC ONLUS Fdn, European Inst Res Cyst Fibrosis, Via Olgettina 58, I-20132 Milan, Italy
[3] Univ Piemonte Orientale, Dept Hlth Sci, I-28100 Novara, Italy
基金
欧盟地平线“2020”;
关键词
three-dimensional microscopy; light sheet fluorescence microscopy; image processing; PLANE ILLUMINATION MICROSCOPY; OPTICAL PROJECTION TOMOGRAPHY; 3-DIMENSIONAL VISUALIZATION; IN-VIVO; ULTRAMICROSCOPY; RESOLUTION; DEEP;
D O I
10.1117/1.JBO.21.5.056001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Light sheet fluorescence microscopy has proven to be a powerful tool to image fixed and chemically cleared samples, providing in depth and high resolution reconstructions of intact mouse organs. We applied light sheet microscopy to image the mouse intestine. We found that large portions of the sample can be readily visualized, assessing the organ status and highlighting the presence of regions with impaired morphology. Yet, three-dimensional (3-D) sectioning of the intestine leads to a large dataset that produces unnecessary storage and processing overload. We developed a routine that extracts the relevant information from a large image stack and provides quantitative analysis of the intestine morphology. This result was achieved by a three step procedure consisting of: (1) virtually unfold the 3-D reconstruction of the intestine; (2) observe it layer-by-layer; and (3) identify distinct villi and statistically analyze multiple samples belonging to different intestinal regions. Even if the procedure has been developed for the murine intestine, most of the underlying concepts have a general applicability. (C) The Authors.
引用
收藏
页数:8
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