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成像形态发生

Imaging morphogenesis.

作者信息

Bell Donald M

机构信息

The Francis Crick Institute Mill Hill Laboratories, The Ridgeway, Mill Hill, London NW7 1AA, UK

出版信息

Philos Trans R Soc Lond B Biol Sci. 2017 May 19;372(1720). doi: 10.1098/rstb.2015.0511.

Abstract

The hostile environment of the microscope stage poses numerous challenges to successful imaging of morphogenesis in live tissues. This review aims to highlight some of the main practical considerations to take into account when embarking on a project to image cell behaviour in the context of cells' normal surroundings. Scrutiny of these activities is likely to be the most informative approach to understanding mechanical morphogenesis but is often confounded by the substantial technical difficulties involved in imaging samples over extended periods of time. Repeated observation of cells in live tissue requires that strategies be adopted to prioritize the stability of the sample, ensuring that it remains viable and develops normally while being held in a manner accessible to microscopic examination. Key considerations when creating reliable protocols for time-lapse imaging may be broken down into three main criteria; labelling, mounting and image acquisition. Choices and compromises made here, however, will directly influence image quality, and even small refinements can substantially improve what information may be extracted from images. Live imaging of tissue is difficult but paying close attention to the basics along with a little innovation is likely to be well rewarded.This article is part of the themed issue 'Systems morphodynamics: understanding the development of tissue hardware'.

摘要

显微镜载物台的恶劣环境给在活组织中成功成像形态发生带来了诸多挑战。本综述旨在强调在开展一个在细胞正常环境背景下对细胞行为进行成像的项目时,需要考虑的一些主要实际问题。仔细研究这些活动可能是理解机械形态发生最具启发性的方法,但长时间对样本进行成像所涉及的巨大技术困难常常使其变得复杂。要对活组织中的细胞进行反复观察,就需要采取策略来优先保证样本的稳定性,确保其在以便于显微镜检查的方式固定时仍保持活力并正常发育。为延时成像创建可靠方案时的关键考虑因素可分为三个主要标准:标记、固定和图像采集。然而,这里做出的选择和权衡将直接影响图像质量,即使是微小的改进也能大幅提升从图像中提取的信息。组织的实时成像很困难,但密切关注基础要点并稍有创新可能会带来丰厚回报。本文是主题为“系统形态动力学:理解组织硬件的发育”的特刊的一部分。

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