CycleTrak: a novel system for the semi-automated analysis of cell cycle dynamics.
CycleTrak: a novel system for the semi-automated analysis of cell cycle dynamics.
复制标题
CycleTrak：一种用于细胞周期动力学半自动分析的新型系统。
DOI：
10.1016/j.ydbio.2012.02.026
复制
发表时间：
2012
影响因子：
2.7
通讯作者：
Kulesa,PaulM
中科院分区：
文献类型：
--
作者：
Ridenour,DennisA;McKinney,MaryCathleen;Bailey,CalebM;Kulesa,PaulM
在胚胎发生过程中，细胞增殖对组织生长和形成至关重要，但细胞周期进程和细胞位置的动态跟踪是一个具有挑战性的障碍。我们已经开发了一种荧光细胞周期指示器和单细胞分析方法，称为CycleTrak，它允许比现有方法更好的时空分辨率和细胞周期阶段和细胞位置的量化。我们的方法是在现有的傅氏法的基础上发展起来的。CycleTrak使用单一慢病毒载体，整合了mKO2-hCdt1（30/120）和一个核定位的eGFP报告基因。单一载体和核定位荧光信号简化了进入细胞的传递，并允许在单个和亚群细胞中快速，自动的细胞跟踪和细胞周期阶段读出。我们验证了CycleTrak在转移性黑色素瘤细胞中的性能，并在移植后的体外和体内鉴定了新的细胞周期动力学，并在活鸡胚胎中进行了3D共聚焦延时成像。
Cell proliferation is crucial to tissue growth and form during embryogenesis, yet dynamic tracking of cell cycle progression and cell position presents a challenging roadblock. We have developed a fluorescent cell cycle indicator and single cell analysis method, called CycleTrak, which allows for better spatiotemporal resolution and quantification of cell cycle phase and cell position than current methods. Our method was developed on the basis of the existing Fucci method. CycleTrak uses a single lentiviral vector that integrates mKO2-hCdt1 (30/120), and a nuclear-localized eGFP reporter. The single vector and nuclear localized fluorescence signals simplify delivery into cells and allow for rapid, automated cell tracking and cell cycle phase readout in single and subpopulations of cells. We validated CycleTrak performance in metastatic melanoma cells and identified novel cell cycle dynamics in vitro and in vivo after transplantation and 3D confocal time-lapse imaging in a living chick embryo.