VBNC细菌荧光观察技术问题分析及对策

Problems Analysis and Solution Strategies for Fluorescence Observation in VBNC Bacteria

摘要:

以SYTO-9和PI两种荧光染料对细菌进行核染, 以市售的黑色钢笔墨水替代伊拉克黑, 利用手动压片方式对标本进行固定, 自建一种“活的非可培养状态”(VBNC)细菌荧光显微镜观察技术。通过总结分析该项技术方法在应用过程中所出现的问题, 如存在背景荧光、菌体聚集、荧光图像模糊、滤膜吸附能力差、菌体形态不鲜明、菌体荧光减淡等, 并提出相应的解决策略, 旨在给予相关研究者以帮助和启迪。

Abstract:

A detection method of viable but non-culturable bacteria by fluorescence microscopy was established with SYTO9 and PI staining cell nucleus, ink instead of Iraq black, specimens fixed by hand. Problems on background fluorescence, cell aggregation, blur fluorescence image, low micropore film absorbability, unclear bacterial form, and fluorescence quenching were analyzed. Some solution strategies were suggested in order to offer help for researchers.