Olson R J, Zettler E R, Anderson O K
Biology Department, Woods Hole Oceanographic Institution, Massachusetts 02543.
Cytometry. 1989 Sep;10(5):636-43. doi: 10.1002/cyto.990100520.
Flow cytometric methods for recognizing several groups of eukaryotic marine phytoplankton were tested using 26 laboratory cultures. Each culture was divided into three aliquots, and these samples were analyzed for 1) Coulter volume; 2) light scatter (magnitude and polarization properties of forward scattered light and magnitude of right-angle scattered light) and autofluorescence emission (phycoerythrin and chlorophyll); and 3) autofluorescence excitation (by 488 nm and 515 nm light). Three kinds of cells could be easily distinguished from others in the culture collection: 1) The two cryptophytes and the rhodophyte had high phycoerythrin/chlorophyll ratios; 2) the two coccolithophores depolarized forward scattered light; and 3) the two pennate diatoms scattered only a relatively small amount of light in the forward direction compared with that at right angles. Mean chlorophyll fluorescence excited by blue light relative to that excited by green light was highest in the four chlorophytes, but there was overlap between some of these and some other kinds of cells. Unresolved cell types included centric diatoms, dinoflagellates, and naked coccolithophores. Forward light scatter and Coulter volume were closely related (except for the pennate diatoms) over a range of about 0.01 to 30 pL (equivalent spherical diameter about 3 to 40 microns), according to a logarithmic function.
使用26种实验室培养物对识别几类真核海洋浮游植物的流式细胞术方法进行了测试。每种培养物被分成三份等分试样，对这些样品进行了以下分析：1）库尔特体积；2）光散射（前向散射光的强度和偏振特性以及直角散射光的强度）和自发荧光发射（藻红蛋白和叶绿素）；3）自发荧光激发（用488nm和515nm光）。在培养物集合中，三种细胞可以很容易地与其他细胞区分开来：1）两种隐藻和红藻具有高藻红蛋白/叶绿素比率；2）两种颗石藻使前向散射光去偏振；3）与直角方向相比，两种羽纹硅藻在前向方向仅散射相对少量的光。蓝光激发的平均叶绿素荧光相对于绿光激发的平均叶绿素荧光在四种绿藻中最高，但其中一些与其他一些细胞类型之间存在重叠。未分辨的细胞类型包括中心硅藻、甲藻和裸颗石藻。根据对数函数，在前向光散射和库尔特体积在约0.01至30皮升（等效球形直径约3至40微米）的范围内密切相关（羽纹硅藻除外）。
