ABSTRACT
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.
Subject(s)
Blastomyces/genetics , Gene Expression Regulation, Fungal , Genes, Reporter , Leukocytes/microbiology , Luminescent Proteins/genetics , Lung/microbiology , Microscopy, Fluorescence , Animals , Blastomyces/immunology , Blastomyces/metabolism , Flow Cytometry , Host-Pathogen Interactions , Humans , Leukocytes/immunology , Leukocytes/metabolism , Luminescent Proteins/biosynthesis , Lung/immunology , Lung/metabolism , Microbial Viability , Red Fluorescent ProteinABSTRACT
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.
