姐妹染色单体交换分析。

Analysis for Sister Chromatid Exchange.

机构信息

Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.

出版信息

Methods Mol Biol. 2023;2519:73-82. doi: 10.1007/978-1-0716-2433-3_8.

Abstract

Sister chromatid exchange (SCE) is the exchange event of genetic material between two identical sister chromatid. Elevation of SCE frequency is considered as a result of replication stress from genetic defects, ROS stress, and genomic damages. SCE staining needs extra processes compared to regular Giemsa staining. Usually two rounds of cell cycle progress are required to observe SCE under microscope. SCE can be visualized with the fluorescence plus Giemsa (FPG) staining method or fluorescence staining methods with immunocytochemistry to BrdU or Click reaction to EdU which provide more clear images of SCE. This chapter will provide the detailed method for the SCE staining and measurement for the traditional FPG staining, BrdU monoclonal antibody staining method, and newly developed EdU Click reaction staining method.

摘要

姐妹染色单体交换（SCE）是两条相同姐妹染色单体之间遗传物质的交换事件。SCE 频率的升高被认为是遗传缺陷、ROS 应激和基因组损伤引起的复制应激的结果。与常规吉姆萨染色相比，SCE 染色需要额外的过程。通常需要两轮细胞周期进展才能在显微镜下观察到 SCE。可以使用荧光加吉姆萨（FPG）染色法或荧光染色方法与 BrdU 或点击反应到 EdU 的免疫细胞化学来可视化 SCE，这为 SCE 提供了更清晰的图像。本章将提供传统 FPG 染色、BrdU 单克隆抗体染色方法和新开发的 EdU 点击反应染色方法的 SCE 染色和测量的详细方法。