Infectious Disease Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
The Robert H. Smith Faculty of Agricultural, Food & Environment, The Hebrew University of Jerusalem, Rehovot, Israel.
Methods Mol Biol. 2021;2260:121-132. doi: 10.1007/978-1-0716-1182-1_8.
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 filamentous mold Aspergillus fumigatus.
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时,以单事件分辨率监测生理过程,特别是真菌细胞的活力和死亡。通过在真菌生物体内预先或在小鼠或培养的白细胞中进行实验感染时加入两种独立的荧光探针,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。本章综述了荧光真菌活力报告器的技术和应用,重点介绍了丝状霉菌烟曲霉。