流式细胞术(FCM)是目前最广泛使用的建立植物核DNA含量的方法。由于简单,1-3-参数,流式细胞仪,这足以满足大多数植物的应用,在商业上以合理的价格购买,配备这些仪器的实验室数量,因此,新的FCM用户,在过去十年中大幅增加。本文满足了针对不同植物科学应用的FCM最佳实践的全面建议的迫切需要。我们讨论了建立植物倍性的优点和局限性,基因组大小,DNA碱基组成,细胞周期活动,和内复制水平。此类测量在植物系统学中的应用,生态学,分子生物学研究,繁殖生物学,组织培养,植物育种,和种子科学的描述。建议包括如何获得准确可靠的结果,以及如何管理样品制备过程中可能发生的故障排除,细胞计数测量,和数据处理。每个部分之后都有最佳实践建议;还提供了有关FCM论文中应提供哪些具体信息的提示。
Flow cytometry (FCM) is currently the most widely-used method to establish nuclear DNA content in plants. Since simple, 1-3-parameter, flow cytometers, which are sufficient for most plant applications, are commercially available at a reasonable price, the number of laboratories equipped with these instruments, and consequently new FCM users, has greatly increased over the last decade. This paper meets an urgent need for comprehensive recommendations for best practices in FCM for different plant science applications. We discuss advantages and limitations of establishing plant ploidy, genome size, DNA base composition, cell cycle activity, and level of endoreduplication. Applications of such measurements in plant systematics, ecology, molecular biology research, reproduction biology, tissue cultures, plant breeding, and seed sciences are described. Advice is included on how to obtain accurate and reliable results, as well as how to manage troubleshooting that may occur during sample preparation, cytometric measurements, and data handling. Each section is followed by best practice recommendations; tips as to what specific information should be provided in FCM papers are also provided.