Longin Cédric, Petitgonnet Clément, Guilloux-Benatier Michèle, Rousseaux Sandrine, Alexandre Hervé
Univ. Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, F-21000 Dijon, France; Institut Universitaire de la Vigne et du Vin, Equipe VAlMiS, rue Claude Ladrey, BP 27877, F-21078 Dijon, France.
Food Microbiol. 2017 Apr;62:221-231. doi: 10.1016/j.fm.2016.10.023. Epub 2016 Oct 18.
Flow cytometry (FCM) is a powerful technique allowing detection and enumeration of microbial populations in food and during food process. Thanks to the fluorescent dyes used and specific probes, FCM provides information about cell physiological state and allows enumeration of a microorganism in a mixed culture. Thus, this technique is increasingly used to quantify pathogen, spoilage microorganisms and microorganisms of interest. Since one decade, FCM applications to the wine field increase greatly to determine population and physiological state of microorganisms performing alcoholic and malolactic fermentations. Wine spoilage microorganisms were also studied. In this review we briefly describe FCM principles. Next, a deep revision concerning enumeration of wine microorganisms by FCM is presented including the fluorescent dyes used and techniques allowing a yeast and bacteria species specific enumeration. Then, the last chapter is dedicated to fluorescent dyes which are used to date in fluorescent microscopy but applicable in FCM. This chapter also describes other interesting "future" techniques which could be applied to study the wine microorganisms. Thus, this review seeks to highlight the main advantages of the flow cytometry applied to wine microbiology.
流式细胞术（FCM）是一项强大的技术，可用于检测和计数食品及食品加工过程中的微生物群体。借助所使用的荧光染料和特异性探针，FCM能够提供有关细胞生理状态的信息，并能对混合培养物中的微生物进行计数。因此，这项技术越来越多地用于量化病原体、腐败微生物以及目标微生物。近十年来，FCM在葡萄酒领域的应用大幅增加，用于确定进行酒精发酵和苹果酸-乳酸发酵的微生物群体及生理状态。同时也对葡萄酒腐败微生物进行了研究。在本综述中，我们简要描述了FCM的原理。接下来，对通过FCM计数葡萄酒微生物进行了深入综述，包括所使用的荧光染料以及能够对酵母和细菌进行特异性计数的技术。最后一章专门介绍了迄今为止用于荧光显微镜但也适用于FCM的荧光染料。本章还描述了其他可能应用于研究葡萄酒微生物的有趣的“未来”技术。因此，本综述旨在突出流式细胞术应用于葡萄酒微生物学的主要优势。
