Imaging embryonic development in Caenorhabditis elegans.
Imaging embryonic development in Caenorhabditis elegans.
复制标题
秀丽隐杆线虫胚胎发育成像。
DOI：
10.1101/pdb.top71
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发表时间：
2010
影响因子：
--
通讯作者：
Isaacson,ArielB
中科院分区：
文献类型：
--
作者：
Mohler,WilliamA;Isaacson,ArielB
胚胎因其多能性、三维性以及亚细胞和发育重排的快速性而引人注目。线虫的胚胎发生在模型系统中特别适合高分辨率动态成像。在单个高放大倍率、高数值孔径 (NA) 显微镜视野内，以亚微米分辨率，可以观察到几只动物的完整形态。这种透明、自由生活的蠕虫的胚胎分裂和形态发生的整个~14小时过程基本上是不变的。在胚胎发育过程中观察特定的荧光标记成分有望揭示细胞器和分子在极其多样化和可重复的环境中的作用。秀丽隐杆线虫社区已经创建了数百种转基因菌株，它们的数量不断增长，这些菌株表达绿色荧光蛋白（GFP）标记的不同内源表达基因。将由此产生的表达和定位模式在空间和时间上关联起来的任务既诱人又技术要求高。本文描述了使用四维 (4D) 激光扫描显微镜和后续数据处理来记录、描绘、分析和比较线虫胚胎发育过程中荧光标记基因产物的表达。
Embryos are remarkable for their combination of pluripotency, three-dimensionality, and swiftness of subcellular and developmental rearrangements. Embryogenesis in the nematode Caenorhabditis elegans is uniquely suited among model systems to high-resolution dynamic imaging. Within a single high-magnification, high-numerical aperture (NA) microscope field, at submicrometer resolution, it is possible to observe several entire animals taking form. The full~ 14-h course of embryonic cleavage and morphogenesis of this transparent, free-living worm is essentially invariant. Observing specific fluorescently labeled components during embryonic development promises to reveal the roles of organelles and molecules in an extremely diverse and reproducible set of contexts. The C. elegans community has created a growing collection of hundreds of transgenic strains expressing green fluorescent protein (GFP)-labeled versions of distinct endogenously expressed genes. The task of correlating the resulting expression and localization patterns in space and time is simultaneously alluring and technically demanding. This article describes the use of four-dimensional (4D) laser-scanning microscopy and subsequent data processing to record, portray, analyze, and compare the expression of fluorescently tagged gene products during development of the nematode embryo.