利用生物荧光技术在小鼠感染模型中可视化缺氧情况。
Visualizing Hypoxia in a Murine Model of Infection Using Biofluorencence.
作者信息
Lopes José Pedro, Urban Constantin F
机构信息
Department of Clinical Microbiology, Umeå University, Umeå, Sweden.
Umeå Centre for Microbial Research, Umeå, Sweden.
出版信息
Bio Protoc. 2019 Aug 5;9(15):e3326. doi: 10.21769/BioProtoc.3326.
Abstract
is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with in a murine infection model. Hypoxia was measured using a fluorescent dye for carbonic anhydrase 9, a host enzyme active under hypoxic conditions. Emitted fluorescence was subsequently quantified using an IVIS system. This protocol was optimized for the use in subdermal infection in mice but has the potential to be adapted to other models of fungal infection.
摘要
是一种主要的人类真菌病原体，它利用多种代谢适应性来逃避免疫细胞并引发全身性疾病。在此，我们描述了一种用于测量其中一种适应性的方案，即在低氧微环境中茁壮成长的能力。在小鼠感染模型中成功进行皮下感染后产生低氧状态。使用针对碳酸酐酶9的荧光染料来测量低氧状态，碳酸酐酶9是一种在低氧条件下具有活性的宿主酶。随后使用IVIS系统对发出的荧光进行定量。该方案针对小鼠皮下感染进行了优化，但有可能适用于其他真菌感染模型。
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