División de Genética, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social, Monterrey, México (EICG,MIDR).
J Histochem Cytochem. 2011 Jul;59(7):655-60. doi: 10.1369/0022155411410884. Epub 2011 May 3.
The comet assay is a well-established, simple, versatile, visual, rapid, and sensitive tool used extensively to assess DNA damage and DNA repair quantitatively and qualitatively in single cells. The comet assay is most frequently used to analyze white blood cells or lymphocytes in human biomonitoring studies, although other cell types have been examined, including buccal, nasal, epithelial, and placental cells and even spermatozoa. This study was conducted to design a protocol that can be used to generate comets in subnuclear units, such as chromosomes. The new technique is based on the chromosome isolation protocols currently used for whole chromosome mounting in electron microscopy, coupled to the alkaline variant of the comet assay, to detect DNA damage. The results show that migrant DNA fragments can be visualized in whole nuclei and isolated chromosomes and that they exhibit patterns of DNA migration that depend on the level of DNA damage produced. This protocol has great potential for the highly reproducible study of DNA damage and repair in specific chromosomal domains.
彗星实验是一种成熟、简单、通用、直观、快速且灵敏的工具，广泛用于定量和定性分析单细胞中的 DNA 损伤和 DNA 修复。彗星实验最常用于分析人类生物监测研究中的白细胞或淋巴细胞，尽管已经检查了其他细胞类型，包括口腔、鼻腔、上皮和胎盘细胞，甚至精子。本研究旨在设计一种可用于在亚核单位（如染色体）中产生彗星的方案。该新技术基于目前用于电子显微镜中整条染色体安装的染色体分离方案，并结合彗星实验的碱性变体，以检测 DNA 损伤。结果表明，可以在整个核和分离的染色体中观察到迁移的 DNA 片段，并且它们表现出的 DNA 迁移模式取决于产生的 DNA 损伤水平。该方案在高度重现性地研究特定染色体区域中的 DNA 损伤和修复方面具有巨大潜力。
