Automated cell lineage tracing in Caenorhabditis elegans
Automated cell lineage tracing in Caenorhabditis elegans
复制标题
DOI：
10.1073/pnas.0511111103
复制
发表时间：
2006-02-21
影响因子：
11.1
通讯作者：
Waterston, RH
中科院分区：
文献类型：
--
作者：
Bao, ZR;Murray, JI;Waterston, RH
秀丽隐杆线虫不变的细胞谱系和细胞命运为解码动物发育的分子机制提供了独特的机会。为了利用这一机会，我们开发了一种基于3D，延时成像和自动图像分析的秀丽隐杆线虫胚胎发生过程中自动细胞谱系追踪系统，使用普遍表达的组蛋白- gfp融合蛋白标记细胞/细胞核和共聚焦显微镜，成像方案在高空间（31个平面间隔1 μ m）和时间（每分钟）分辨率下捕获胚胎发生，没有明显影响！在发展。然后，一组图像分析算法自动识别每个时间点的细胞，跟踪细胞的运动、分裂和死亡，并根据规范命名方案分配细胞身份。从四细胞阶段（或更早）开始，我们名为STARRYNITE的软件可以在台式计算机上25分钟内追踪到350细胞阶段的谱系。自动化谱系的一些错误可以在几个小时内通过图形界面进行纠正，该界面允许轻松地导航图像和报告的谱系树。该系统可用于表征基因的谱系表型和/或扩展以确定单细胞水平活胚胎中的基因表达模式。我们设想，这种自动化将使系统地破译秀丽隐杆线虫基因组中编码的发育基因和途径成为现实。
The invariant cell lineage and cell fate of Caenorhabditis elegans provide a unique opportunity to decode the molecular mechanisms of animal development. To exploit this opportunity, we have developed a system for automated cell lineage tracing during C elegans embryogenesis, based on 3D, time-lapse imaging and automated image analysis, Using ubiquitously expressed histone-GFP fusion protein to label cells/nuclei and a confocal microscope, the imaging protocol captures embryogenesis at high spatial (31 planes at 1 mu m apart) and temporal (every minute) resolution without apparent effect! on development. A set of image analysis algorithms then automatically recognizes cells at each time point, tracks cell movements, divisions and deaths over time and assigns cell identities based on the canonical naming scheme. Starting from the four-cell stage (or earlier), our software, named STARRYNITE, can trace the lineage up to the 350-cell stage in 25 min on a desktop computer. The few errors of automated lineaging can then be corrected in a few hours with a graphic interface that allows easy navigation of the images and the reported lineage tree. The system can be used to characterize lineage phenotypes of genes and/or extended to determine gene expression patterns in a living embryo at the single-cell level. We envision that this automation will make it practical to systematically decipher the developmental genes and pathways encoded in the genome of C elegans.