DOUBLE IMMUNOFLUORESCENCE MICROSCOPIC TECHNIQUE FOR ACCURATE DIFFERENTIATION OF EXTRACELLULARLY AND INTRACELLULARLY LOCATED BACTERIA IN CELL-CULTURE
DOUBLE IMMUNOFLUORESCENCE MICROSCOPIC TECHNIQUE FOR ACCURATE DIFFERENTIATION OF EXTRACELLULARLY AND INTRACELLULARLY LOCATED BACTERIA IN CELL-CULTURE
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DOI：
10.1128/jcm.22.2.168-175.1985
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发表时间：
1985-01-01
影响因子：
9.4
通讯作者：
LAUFS, R
中科院分区：
文献类型：
--
作者：
HEESEMANN, J;LAUFS, R
活的哺乳动物细胞的膜对抗体是不可渗透的，但通过用固定剂处理而变得可渗透。因此，细胞外细菌可以在固定前被特异性罗丹明标记的抗体染色，而细胞内细菌可以在固定后通过用特异性荧光素标记的抗体处理而可视化。证明了该方法的准确性和简单性与HEp-2细胞培养单层作为靶细胞和小肠结肠炎耶尔森氏菌的同基因对，其中之一是吞噬抗性，其中之一是吞噬敏感。这种染色技术也适用于研究细菌与巨噬细胞和成纤维细胞的相互作用。
Membranes of viable mammalian cells are impermeable to antibodies but are rendered permeable by treatment with fixatives. Consequently, extracellular bacteria can be stained by specific rhodamine-labeled antibodies before fixation, and intracellular bacteria can be visualized by treatment with specific-fluorescein-labeled antibodies after fixation. The accuracy and simplicity of this method was demonstrated with HEp-2 cell culture monolayers as target cells and an isogenic pair of Yersinia enterocolitica, one of which is phagocytosis-resistant and the other of which is phagocytosis-sensitive. This staining technique ia alsao applicable for studying the interaction of bacteria with macrophages and fibroblasts.