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用3',6'-二乙酰荧光素对微生物进行活体荧光染色以测定其代谢活性(作者译)

[Vital fluorescent staining of microorganisms by 3',6'-diacetyl-fluoresceine for determination of their metabolic activity (author's transl)].

作者信息

Ziegler G B, Ziegler E, Witzenhausen R

出版信息

Zentralbl Bakteriol Orig A. 1975;230(2):252-64.

PMID:49989
Abstract

By fluorescent microscopy and spectroscopy studies involving micro-organisms which were either viable or devitalised by heat sterilisation or gamma irradiation and could not be cultured any more, fluorochrome binding with 3' , 6'-diacetyl fluorescein was shown to be linked with the viability of a cell and a function of its actual metabolic state. The incorporation of diacetyl fluorescein into cells, its storage and hydrolysis to fluorescein mean active processes taking place at high speed. Viable cells are capable of storing fluorescein intracellularly, bound to structural elements. If the storage capacity is surpassed, it will be eliminated from the cell. The mechanism of this process is discussed. Devitalised cells are not capable of active uptake of fluorescein nor of its storage and accelerated hydrolysis. Beyond this, they are incapable of fluorescein binding to structural elements. There will be only a minor homogenous staining of such cells by fluorescein. An express method based upon the results is providing information on viability, actual metabolic state, morphology, and motility of micro-organisms within a few minutes and without affecting onward culturing.

摘要

通过荧光显微镜和光谱学研究,涉及经热灭菌或伽马射线辐照而存活或失活且无法再培养的微生物,结果表明,与3',6'-二乙酰荧光素结合的荧光染料与细胞活力及其实际代谢状态的功能有关。二乙酰荧光素掺入细胞、储存并水解为荧光素意味着高速发生的活跃过程。活细胞能够在细胞内储存荧光素,并与结构成分结合。如果储存能力超过限度,荧光素将被排出细胞。本文讨论了这一过程的机制。失活细胞既不能主动摄取荧光素,也不能储存和加速水解荧光素。此外,它们无法使荧光素与结构成分结合。此类细胞只会被荧光素轻微均匀染色。基于这些结果的一种快速方法能够在几分钟内提供有关微生物活力、实际代谢状态、形态和运动性的信息,且不会影响后续培养。

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