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Author:
Date:
2016-07-20
[Abstract] Nitric Oxide (NO) is a highly-reactive radical gas that can modify a variety of cellular targets in both eukaryotes and bacteria. NO is produced endogenously by a wide variety of organisms: For example, as a cell-signaling molecule in mammals and bacteria via nitric oxide synthase (NOS) enzymes, and as a product of denitrification. As such, it is of great benefit to NO researchers to be able to sensitively detect intracellular NO and stable reactive nitrogen species (RNS) derived from NO. To this end, a protocol for fluorescent detection of intracellular NO/RNS in biofilm cultures of the Gram-positive pathogen Staphylococcus aureus has been optimized using the commercially-available cell-permeable fluorescent stain 4-Amino-5-Methylamino-2’,7’-Difluorofluorescein Diacetate (DAF-FM ...
[摘要] 一氧化氮（NO）是一种高反应性的自由基气体，其可以修饰真核生物和细菌中的多种细胞靶标。 NO由多种生物体内源性产生：例如，作为哺乳动物和细菌中的细胞信号分子，通过一氧化氮合酶（NOS）酶，以及作为脱氮的产物。因此，NO研究人员能够敏感地检测来自NO的细胞内NO和稳定的活性氮物质（RNS）是非常有益的。为此，已经使用商业上可获得的细胞可渗透的荧光染料4-氨基-5来优化用于荧光检测革兰氏阳性病原体金黄色葡萄球菌的生物膜培养物中的细胞内NO/RNS的方案 - 甲基氨基-2'，7'-二氟荧光素二乙酸酯（DAF-FM二乙酸酯）。该化合物扩散到细胞中并且通过酯酶的细胞内裂解释放弱荧光DAF-FM，其与NO或其它特异性RNS反应以变成高度荧光的（Kojima等人，1999）。虽然使用荧光板读数器进行荧光的定量，但设想该方案可适用于S的细胞内NO/RNS成像。 aureus biofilms by confocal microscopy。同样，这种技术可以被优化用于检测其它生长条件（即浮游生物培养物）和/或其它细菌/古细菌中的细胞内NO/RNS。
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