Department of Pediatrics, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, USA.
Fungal Pathogenesis Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2021;2260:111-119. doi: 10.1007/978-1-0716-1182-1_7.
Fluorescence-based techniques enable researchers to monitor physiologic processes, specifically fungal cell viability and death, during cellular encounters with the mammalian immune system with single event resolution. By incorporating two independent fluorescent probes in fungal organisms either prior to, or ensuing experimental infection in mice or in cultured leukocytes, it is possible to distinguish and quantify live and killed fungal cells to interrogate genetic, pharmacologic, and cellular determinants that shape host-fungal cell outcomes. This chapter reviews the techniques and applications of fluorescent fungal reporters of viability, with emphasis on the North American endemic dimorphic fungus, Blastomyces dermatitidis.
基于荧光的技术使研究人员能够在哺乳动物免疫系统与细胞相互作用的过程中，以单细胞分辨率监测生理过程，特别是真菌细胞的活力和死亡。通过在真菌生物中预先或在小鼠或培养的白细胞中进行实验感染时，引入两种独立的荧光探针，就可以区分和量化活细胞和死细胞，从而研究影响宿主-真菌细胞结果的遗传、药理学和细胞决定因素。本章回顾了荧光真菌生存力报告器的技术和应用，重点介绍了北美地方性二相真菌——皮炎芽生菌。
