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利用黄素相关细胞自发荧光监测微生物生物技术中的过程。

Use of Flavin-Related Cellular Autofluorescence to Monitor Processes in Microbial Biotechnology.

作者信息

Müllerová Lucie, Marková Kateřina, Obruča Stanislav, Mravec Filip

机构信息

Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic.

出版信息

Microorganisms. 2022 Jun 8;10(6):1179. doi: 10.3390/microorganisms10061179.

Abstract

Cellular autofluorescence is usually considered to be a negative phenomenon because it can affect the sensitivity of fluorescence microscopic or flow cytometric assays by interfering with the signal of various fluorescent probes. Nevertheless, in our work, we adopted a different approach, and green autofluorescence induced by flavins was used as a tool to monitor fermentation employing the bacterium . The autofluorescence was used to distinguish microbial cells from abiotic particles in flow cytometry assays, and it was also used for the determination of viability or metabolic characteristics of the microbial cells. The analyses using two complementary techniques, namely fluorescence microscopy and flow cytometry, are simple and do not require labor sample preparation. Flavins and their autofluorescence can also be used in a combination with other fluorophores when the need for multi-parametrical analyses arises, but it is wise to use dyes that do not emit a green light in order to not interfere with flavins' emission band (500-550 nm).

摘要

细胞自发荧光通常被视为一种负面现象,因为它会干扰各种荧光探针的信号,从而影响荧光显微镜或流式细胞术检测的灵敏度。然而,在我们的工作中,我们采用了不同的方法,利用黄素诱导的绿色自发荧光作为监测该细菌发酵的工具。在流式细胞术检测中,自发荧光被用于区分微生物细胞和非生物颗粒,还用于测定微生物细胞的活力或代谢特征。使用荧光显微镜和流式细胞术这两种互补技术进行分析很简单,且无需繁琐的样品制备。当需要进行多参数分析时,黄素及其自发荧光也可与其他荧光团结合使用,但明智的做法是使用不发射绿光的染料,以免干扰黄素的发射带(500 - 550纳米)。

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