An Updated Protocol for High Throughput Plant Tissue Sectioning.
An Updated Protocol for High Throughput Plant Tissue Sectioning.
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DOI：
10.3389/fpls.2017.01721
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发表时间：
2017
影响因子：
5.6
通讯作者：
Wells DM
中科院分区：
文献类型：
--
作者：
Atkinson JA;Wells DM
植物材料的组织和细胞结构的量化对于研究各种植物科学应用至关重要。目前，许多植物材料切片方法要么产量低，要么需要徒手切片，这需要大量的练习。在这里，我们提出了一种更新的方法，可以提供快速且高质量的横截面，主要是根组织的横截面，但也可以轻松应用于其他组织，例如叶子或茎。为了提高传统琼脂糖嵌入和切片的吞吐量，使用定制设计的 3D 打印模具将 5-15 个根嵌入块中，以便在单次切割中进行切片。使用单一荧光染色与激光扫描共焦显微镜相结合来获得厚切片的高质量图像。所提供的 CAD 文件允许通过许多在线 3D 打印服务生产此处描述的嵌入模具。尽管该方法最初是针对根部开发的，但它提供了许多植物组织类型的快速、高质量的横截面，使其适合用于特定细胞结构差异或发育变化的正向遗传筛选。为了证明该技术的实用性，对中国春×Paragon双单倍体作图群体的两个亲本系的根解剖差异进行了表型分析。发现不定横截面积、中柱面积、木质部、韧皮部、后木质部和皮质细胞锉计数存在显着差异。
Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practice. Here, we present an updated method to provide rapid and high-quality cross sections, primarily of root tissue but which can also be readily applied to other tissues such as leaves or stems. To increase the throughput of traditional agarose embedding and sectioning, custom designed 3D printed molds were utilized to embed 5–15 roots in a block for sectioning in a single cut. A single fluorescent stain in combination with laser scanning confocal microscopy was used to obtain high quality images of thick sections. The provided CAD files allow production of the embedding molds described here from a number of online 3D printing services. Although originally developed for roots, this method provides rapid, high quality cross sections of many plant tissue types, making it suitable for use in forward genetic screens for differences in specific cell structures or developmental changes. To demonstrate the utility of the technique, the two parent lines of the wheat (Triticum aestivum) Chinese Spring × Paragon doubled haploid mapping population were phenotyped for root anatomical differences. Significant differences in adventitious cross section area, stele area, xylem, phloem, metaxylem, and cortical cell file count were found.
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影响因子：
6.9
作者：
Prince SJ;Murphy M;Mutava RN;Durnell LA;Valliyodan B;Shannon JG;Nguyen HT
通讯作者：
Nguyen HT
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作者：
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通讯作者：
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作者：
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通讯作者：
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