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FDA-PI双色荧光法检测蓝藻细胞活性的研究

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谌丽斌, 梁文艳, 曲久辉, 解明曙, 雷鹏举, 刘会娟. FDA-PI双色荧光法检测蓝藻细胞活性的研究[J]. 环境化学, 2005, 24(5): 554-557.
引用本文: 谌丽斌, 梁文艳, 曲久辉, 解明曙, 雷鹏举, 刘会娟. FDA-PI双色荧光法检测蓝藻细胞活性的研究[J]. 环境化学, 2005, 24(5): 554-557.
shu
CHEN Li-bin, LIANG Wen-yan, QU Jiu-hui, XIE Ming-shu, LEI Peng-jiu, LIU Hui-juan. THE VIABILITY DETERMINATION OF CYANOBACTERIA BY DOUBLE STAINING WITH FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE[J]. Environmental Chemistry, 2005, 24(5): 554-557.
Citation: CHEN Li-bin, LIANG Wen-yan, QU Jiu-hui, XIE Ming-shu, LEI Peng-jiu, LIU Hui-juan. THE VIABILITY DETERMINATION OF CYANOBACTERIA BY DOUBLE STAINING WITH FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE[J]. Environmental Chemistry, 2005, 24(5): 554-557.
shu

FDA-PI双色荧光法检测蓝藻细胞活性的研究

  • 1.

    中国科学院生态环境研究中心, 环境水质学国家重点实验室, 北京, 100085;

  • 2.

    北京林业大学资源与环境学院, 北京, 100083

  • 基金项目:

    国家自然科学基金(50478115)

    国家自然科学基金重点基金(20337020)

THE VIABILITY DETERMINATION OF CYANOBACTERIA BY DOUBLE STAINING WITH FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE

  • 1.

    State Key Laboratory of Environmental Aquatic Chemistry, Res earchCenter for Eco-Environmental Science, Chinese Academy of Science, Beijin g, 100085;

  • 2.

    College of Natural Resources & Environment, Beijing Forestry University, Beijing, 100083

  • Fund Project:

  • 摘要: 对FDA-PI双色荧光法检测水华鱼腥藻和铜绿微囊藻的活性进行了研究,用荧光显微镜对染色结果进行测定.结果表明,在蓝色光激发下(495nm),活细胞被双醋酸荧光素FDA染成亮绿色,死亡细胞被碘化丙锭PI染成红色.染色效率与原植体类型和细胞密度有关.对细胞密度为6×107—7×109个·l-1的铜绿微囊藻,FDA染色效率可达94%以上.对细胞密度为4×107—5×108个·l-1的水华鱼腥藻,FDA染色效率可达91%以上,但细胞密度增大到5×109个·l-1时,由于藻丝体易卷曲在一起,FDA染色率下降到67%.对死亡细胞,PI染色率基本都可达到100%.因此,用FDA-PI检测活细胞和死亡细胞混合的细胞悬液,可根据细胞所发出的不同荧光而判断细胞活性.
    Abstract: The paper studied the viability determination of two kinds of algae Anabae na flos-aquae and Microcystis aeruginosa by FDA-PIdouble staining. Fluo rescence emission from cells was measured by fluorescence microscopy. The result s indicated that viable and dead cells were stained as bright green and red fluo rescent cells respectively by fluorescein diacetate and propidium iodide under b lue excitation(495nm). The staining rate has relation with the cell density. If the cell density of Anabaena flos-aquae and Microcystis aeruginosa was 6×107—7×109cells·l-1 and 4×107—5×108cel ls·l-1, the FDAstaining rate would be more than 94% and 91%. But if the cell density of Anabaena flos-aquae increased to 5×109 cells·l-1,the staining rate would fall to 67% obviously due to cell twisting togethe r. The staining rate of PI can reach 100% for the two kinds of algae. Based on t he fluorescent light from cells, it can be easy to determinate the cell viabilit y. FDA-PI double staining method is a new method to measure the algae cell viability.
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  • [1] 吴星五,高延耀,李国建,电化学法水处理新技术--杀菌灭藻.环境科学学报, 2000,20(增刊):75-79
    [2] Caavate J P, Lubińn L M, Some Aspects on the Cryopreservation of Microalgae Used as Food for Marine Species.Aquaculture,1995,136:277-290
    [3] Veal D A, Deere D, Ferrari B et al., Fluorescence Staining and Flow Cytometry for Monitoring Microbial Cells.Journal Immunological Methods,2000,243:191-210
    [4] Regel R H,Ferris J M, Ganf G G et al., Algal Esterase Activity as a Biomeasure of Environmental Degradation in a Freshwater Creek.Aquatic Toxicology,2002,59:209-223
    [5] Jones K H, Senet J A, An Improved Method to Determine Cell Viability by Simultaneous Staining with Fluorescein Diactqate-propidium Iodide.The Journal of Histochemistry and Cytochemistry,1985,33(1):77-79
    [6] Amano T,Hirasawa K,O' Donohue M J et al., A Versatile Assay for the Accurate, Time-resolved Determination of Cellular Viability.Analytical Biochemistry,2003,314:1-7
    [7] 祁爱群,钱凯先,邵健忠等, FDA-PI双色荧光分光光度法检测细胞活性变化.细胞生物学杂志, 2000,22(1):50-52
    [8] 林碧琴,姜彬慧,藻类与环境保护.沈阳:沈阳民族出版,1999,573
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  • 收稿日期:  2004-12-03
谌丽斌, 梁文艳, 曲久辉, 解明曙, 雷鹏举, 刘会娟. FDA-PI双色荧光法检测蓝藻细胞活性的研究[J]. 环境化学, 2005, 24(5): 554-557.
引用本文: 谌丽斌, 梁文艳, 曲久辉, 解明曙, 雷鹏举, 刘会娟. FDA-PI双色荧光法检测蓝藻细胞活性的研究[J]. 环境化学, 2005, 24(5): 554-557.
shu
CHEN Li-bin, LIANG Wen-yan, QU Jiu-hui, XIE Ming-shu, LEI Peng-jiu, LIU Hui-juan. THE VIABILITY DETERMINATION OF CYANOBACTERIA BY DOUBLE STAINING WITH FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE[J]. Environmental Chemistry, 2005, 24(5): 554-557.
Citation: CHEN Li-bin, LIANG Wen-yan, QU Jiu-hui, XIE Ming-shu, LEI Peng-jiu, LIU Hui-juan. THE VIABILITY DETERMINATION OF CYANOBACTERIA BY DOUBLE STAINING WITH FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE[J]. Environmental Chemistry, 2005, 24(5): 554-557.
shu

FDA-PI双色荧光法检测蓝藻细胞活性的研究

  • 1.  中国科学院生态环境研究中心, 环境水质学国家重点实验室, 北京, 100085;
  • 2.  北京林业大学资源与环境学院, 北京, 100083
  • 收稿日期:  2004-12-03
基金项目:

国家自然科学基金(50478115)

国家自然科学基金重点基金(20337020)

摘要: 对FDA-PI双色荧光法检测水华鱼腥藻和铜绿微囊藻的活性进行了研究,用荧光显微镜对染色结果进行测定.结果表明,在蓝色光激发下(495nm),活细胞被双醋酸荧光素FDA染成亮绿色,死亡细胞被碘化丙锭PI染成红色.染色效率与原植体类型和细胞密度有关.对细胞密度为6×107—7×109个·l-1的铜绿微囊藻,FDA染色效率可达94%以上.对细胞密度为4×107—5×108个·l-1的水华鱼腥藻,FDA染色效率可达91%以上,但细胞密度增大到5×109个·l-1时,由于藻丝体易卷曲在一起,FDA染色率下降到67%.对死亡细胞,PI染色率基本都可达到100%.因此,用FDA-PI检测活细胞和死亡细胞混合的细胞悬液,可根据细胞所发出的不同荧光而判断细胞活性.

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