玉米分生组织的新鲜组织扫描电镜(SEM)。

Comfort Bonney Arku, Madelaine Bartlett, Annis E Richardson
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引用次数: 0

摘要

在植物中,最终器官的形状和位置影响着整个植物的功能和生产力。在玉米中,叶片、穗和雄穗形态都是影响产量的关键农艺特征。这些空中结构来源于分生组织内的干细胞群。了解分生组织是如何调控的,以及干细胞是如何在发育过程中分化形成成熟的叶片、穗和穗的,对于确定玉米改良的新途径非常重要。分生组织形态学的高分辨率成像是发育研究的关键组成部分。扫描电子显微镜(SEM)被广泛用于观察分生组织拓扑结构;然而,传统的SEM制备需要固定、脱水、干燥和导电涂层。这些步骤耗时,技术要求高,并且可能会造成损害。在这里,我们提出了一种使用台式扫描电子显微镜对新鲜解剖的玉米分生组织进行成像的快速方案。新鲜组织直接安装在导电粘合剂上,并在高真空和低加速电压下成像,从而在解剖后几分钟内捕获高质量的显微照片。尽管在真空条件下会发生脱水和组织塌陷,但快速的样品处理和图像采集可以最大限度地减少这些影响,并产生与常规制备标本相当的图像。这种流线型的工作流程大大减少了准备时间,降低了材料要求,并促进了中等通量形态分析。该方法为快速发育表型分析提供了一种实用且可重复的方法,并且易于适用于其他植物物种和其他组织。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Fresh Tissue Scanning Electron Microscopy (SEM) of Maize Meristems.

In plants, final organ shape and position influence function and whole plant productivity. In Zea mays (maize), leaf, ear, and tassel morphology are all key agronomic features influencing yield. These aerial structures originate from stem cell populations within meristems. Understanding how meristems are regulated and how stem cells differentiate over developmental time to form mature leaves, ears, and tassels is important for identifying novel routes for maize improvement. High-resolution imaging of meristem morphology is a key component of developmental studies. Scanning electron microscopy (SEM) is widely used to visualize meristem topology; however, conventional SEM preparation requires fixation, dehydration, drying, and conductive coating. These steps are time-consuming, technically demanding, and can introduce damage. Here, we present a rapid protocol for imaging freshly dissected meristems from maize using a benchtop scanning electron microscope. Fresh tissues are mounted directly onto conductive adhesive and imaged under high vacuum at low accelerating voltage, enabling high-quality micrographs to be captured within minutes of dissection. Although dehydration and tissue collapse can occur under vacuum, rapid sample handling and image acquisition minimize these effects and produce images comparable to those obtained from conventionally prepared specimens. This streamlined workflow substantially reduces preparation time, lowers material requirements, and facilitates medium-throughput morphological analysis. The method provides a practical and reproducible approach for rapid developmental phenotyping and is readily adaptable to other plant species and other tissues.

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来源期刊
Cold Spring Harbor protocols
Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.00
自引率
0.00%
发文量
163
期刊介绍: Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.
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