Ercolini Danilo, Hill Philip J, Dodd Christine E R
Division of Food Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE125RD, UK.
J Microbiol Methods. 2003 Feb;52(2):267-71. doi: 10.1016/s0167-7012(02)00162-8.
A 16S rRNA fluorescence in situ hybridization (FISH) method for cheese was developed to allow detection in situ of microorganisms within the dairy matrix. An embedding procedure using a plastic resin was applied to Stilton cheese, providing intact embedded cheese sections withstanding the hybridization reaction. The use of a fluorescein-labelled 16S rRNA Domain Bacteria probe allowed observation of large colonies of microbial cells homogeneously distributed in the cheese matrix. FISH experiments performed on cheese suspensions provided images of the different microbial morphotypes occurring. The technique has great potential to study the spatial distribution of microbial populations in situ in foods, especially where the matrix is too fragile to allow manipulation of cryosections.
开发了一种用于奶酪的16S rRNA荧光原位杂交（FISH）方法，以便在乳制品基质中原位检测微生物。采用塑料树脂包埋程序处理斯第尔顿奶酪，得到的完整包埋奶酪切片能够承受杂交反应。使用荧光素标记的16S rRNA细菌结构域探针，可以观察到均匀分布在奶酪基质中的大量微生物细胞菌落。对奶酪悬浮液进行的FISH实验提供了不同微生物形态类型的图像。该技术在原位研究食品中微生物种群的空间分布方面具有巨大潜力，尤其是在基质过于脆弱而无法进行冷冻切片操作的情况下。
