Phinney D A, Cucci T L
J.J. MacIsaac Flow Cytometry/Cell Sorting Facility, Bigelow Laboratory for Ocean Sciences, McKown Point, West Boothbay Harbor, Maine 04575.
Cytometry. 1989 Sep;10(5):511-21. doi: 10.1002/cyto.990100506.
Flow cytometry and sorting are now an important technology in aquatic research. Simultaneous measurements of individual particle cell size, fluorescence, and light scatter properties are directly applicable to current topics in aquatic research. Flow sorting may be employed to obtain subsets of cells for analysis by conventional methods. The manner in which rapid, precise measurements of single cells are made is complex, and the application of this technology to aquatic samples is subject to many analytical constraints. Flow cytometric measurements of algal cell size and pigment autofluorescence are relative and are therefore dependent on the optical configuration and variability of the instrument. Specific types of reference materials are used to establish the validity of analyses: 1) instrument standards, 2) fluorescence controls, and 3) internal stain standards. The selection and application of standards and controls are discussed in the context of allometric (cell size versus pigment fluorescence) and ataxonomic (pigment color groups) methods. The widespread acceptance of particular reference materials among research groups will result in comparable data sets describing aquatic particle distributions.
流式细胞术和分选技术如今已成为水生研究中的一项重要技术。对单个颗粒的细胞大小、荧光和光散射特性进行同步测量，可直接应用于当前水生研究的相关课题。可采用流式分选来获取细胞亚群，以便通过传统方法进行分析。进行单细胞快速、精确测量的方式较为复杂，且将该技术应用于水生样本时会受到诸多分析限制。藻类细胞大小和色素自发荧光的流式细胞术测量是相对的，因此取决于仪器的光学配置和变异性。使用特定类型的参考材料来确定分析的有效性：1）仪器标准品，2）荧光对照品，3）内部染色标准品。在异速生长（细胞大小与色素荧光）和非分类学（色素颜色组）方法的背景下，讨论了标准品和对照品的选择与应用。特定参考材料在各研究组中得到广泛认可，将产生描述水生颗粒分布的可比数据集。
