Department of Microbiology and Immunology, Center for Human Immunology, The University of Western Ontario, London, ON, Canada.
Robarts Research Institute, London, ON, Canada.
Methods Mol Biol. 2022;2440:99-114. doi: 10.1007/978-1-0716-2051-9_6.
Fluorescent biosensors are chemically or genetically encoded reporters of cellular processes, signaling pathways, or biomolecule concentration, whose output is quantified using fluorescence microscopy or fluorescence spectrometry. These biosensors can detect the target activity or metabolites via mechanisms including conversion between nonfluorescent and fluorescent forms, changes in reporter intensity, changes in the intensity ratio across fluorescence channels, alterations to the subcellular localization of the bioreporter, and by fluorescence resonance energy transfer. Here, we describe the use of a chemical photoconverting biosensor, and genetically encoded localization and ratiometric biosensors, for monitoring the cellular and signaling processes involved in pathogen-induced apoptosis and the resulting destruction of the pathogen. While this study uses biosensors to monitor responses to infection, these approaches can be readily translated to other cellular systems and other fluorescent biosensors.
荧光生物传感器是细胞过程、信号通路或生物分子浓度的化学或基因编码报告器,其输出使用荧光显微镜或荧光光谱法进行定量。这些生物传感器可以通过以下机制检测目标活性或代谢物:包括非荧光和荧光形式之间的转换、报告器强度的变化、荧光通道之间的强度比的变化、生物报告器的亚细胞定位的改变以及荧光共振能量转移。在这里,我们描述了使用化学光转化生物传感器、基因编码定位和比率生物传感器来监测与病原体诱导的细胞凋亡和随后的病原体破坏相关的细胞和信号过程。虽然本研究使用生物传感器来监测对感染的反应,但这些方法可以很容易地转化为其他细胞系统和其他荧光生物传感器。