海洋病毒荧光显微计数法的优化与应用

Rapid epifluorescence enumeration of marine viruses: method optimization and application

摘要:

【目的】建立一种快速、稳定、可靠的海洋病毒计数方法。【方法】海水水样经福尔马林固定后，滤过孔径为0.02 μm的Anodisc Al2O3膜。滤膜经SYBR Green I染色后，在相应波长的激发光下进行观察。借助荧光显微镜目镜网格尺，计数视野中的病毒颗粒，换算后获得样品中病毒的浓度。【结果】对具体实验方法进行了优化，可快速、稳定地对海水中的病毒计数。【结论】建立了一种适用于国内实验条件的、可靠的海洋病毒计数方法。

Abstract:

[Objective] To establish the epifluorescence microscopic (EFM) method for rapid and reliable enumeration of marine viruses. [Methods] Fixed sea water samples were filtered through 0.02 μm filter, and virus particles were collected on the membrane. After stained with SYBR Green I, viruses were observed and counted under epifluorescence microscope using a grid micrometer. [Results] The EFM method was optimized to count marine viruses rapidly and reliably. [Conclusion] The epifluorence enumeration method was established as well as applied to count marine viruses.