Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.
Department of Pediatrics and Human Development & Department of Pharmacology & Toxicology, Michigan State University, East Lansing, MI, USA.
Methods Mol Biol. 2023;2519:17-26. doi: 10.1007/978-1-0716-2433-3_3.
Cellular division is a fundamental process of cellular growth. First, cells replicate their DNA in S phase and then undergo mitosis which, under normal conditions, leads to complete cell division. Moreover, mitotic activity correlates to cellular growth activity. The simplest and classical method to measure mitotic activity (mitotic index (MI)), is the manual counting of mitotic cells among a given cell population of interest. The latter can be accomplished via phase contrast microscope observation. However, Giemsa staining may improve accuracy and consistency. Fluorescence immunostaining targeting specific phosphorylations of proteins at critical cell cycle steps will provide further improved analysis via high-throughput capacity of flow or imaging cytometer. Finally, time lapse image analysis provides quantitative and qualitative metrics delineating the process of cellular division including timing of division, duration of mitosis, and failure to procced through or complete mitosis.
细胞分裂是细胞生长的基本过程。首先，细胞在 S 期复制其 DNA，然后进行有丝分裂，在正常情况下，有丝分裂导致完全细胞分裂。此外，有丝分裂活性与细胞生长活性相关。测量有丝分裂活性（有丝分裂指数（MI））的最简单和经典的方法是手动计数给定细胞群体中的有丝分裂细胞。后者可以通过相差显微镜观察来完成。然而，吉姆萨染色可以提高准确性和一致性。针对细胞周期关键步骤中特定蛋白质磷酸化的荧光免疫染色将通过流式细胞仪或成像细胞仪的高通量能力提供进一步的改进分析。最后，延时图像分析提供了定量和定性的指标，描绘了细胞分裂的过程，包括分裂的时间、有丝分裂的持续时间以及未能通过或完成有丝分裂的情况。
