藻细胞计数及死/活分析的流式细胞仪方法  

Flow Cytometry Protocols on Algal Cell Counting and Live/Dead Assay for Water Supply

作　　者：陈慧婷[1] 陶益[2] 朱佳[3] 

机构地区：[1]深圳市环境工程科学技术中心,广东深圳518055 [2]清华大学深圳研究生院环境工程与管理研究中心,广东深圳518055 [3]深圳职业技术学院建筑与环境工程学院,广东深圳518055

出　　处：《深圳职业技术学院学报》2013年第1期23-27,共5页Journal of Shenzhen Polytechnic

基　　金：国家自然科学基金(编号50978170);公益性行业科研专项(环保行业)(编号201209009);水专项项目(编号2009ZX07423-001);水利部公益性行业专项(201201033)资助项目

摘　　要：以我国原水典型藻类铜绿微囊藻和椭圆小球藻为例，建立了采用双组分绝对计数微球进行藻细胞计数的流式细胞仪分析方法，以及采用染料SYBR green I和碘化丙啶（PI）双染色判定细胞死／活的流式细胞仪分析方法，运用所建立方法开展了0-200 mJ·cm^-2紫外消毒处理后铜绿微囊藻的细胞计数及死／活分析.研究结果表明，流式细胞方法计数结果与显微计数结果具有较好的线性相关性（R0=0．980），采用流式细胞仪使单样品操作时间缩短至5min以内．流式细胞方法能灵敏检测细胞死／活状态，测定梯度配比的热处理致死细胞的结果与配比值良好吻合，铜绿微囊藻拟合曲线尺。值达0．999，椭圆小球藻拟合曲线彤值0．995．流式细胞方法用于紫外消毒处理水样中藻细胞计数及死／活分析灵敏有效．With two typical strains in China waters as studied subjects, flow cytometry protocols for algal cell counting through Caltag counting beads and for algal live/dead assay through SYBR green I and propidium iodide were established by using Microcystis aeruginosa and Chlorella ellipsoidea. The protocols were adopted for cell counting and live/dead assay of the algal cells with UV-C disinfection treatment at 0-200 mJ.cm2. The results indicate that the cell counting data based on flow cytometry analysis were well linear-correlated with that of microscopic analysis （R2 = 0.980） while saving the detection time to less than 5 min. Live/dead assay data based on flow cytometry were well linear-correlated with the mixture setting with R2 of 0.999 for M. aeruginosa and R2 of 0.995 for C. ellipsoidea, indicating the protocol for live/dead assay was precise. The flow cytometric results of cell counting and live/dead assay of the algal samples with UV-C disinfection treatment show that the protocols were accurate and precise.

关 键 词：藻 流式细胞术 细胞计数 细胞死 活分析 

分 类 号：X524[环境科学与工程—环境工程]