RESUMO
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.
Assuntos
Blastomyces/genética , Regulação Fúngica da Expressão Gênica , Genes Reporter , Leucócitos/microbiologia , Proteínas Luminescentes/genética , Pulmão/microbiologia , Microscopia de Fluorescência , Animais , Blastomyces/imunologia , Blastomyces/metabolismo , Citometria de Fluxo , Interações Hospedeiro-Patógeno , Humanos , Leucócitos/imunologia , Leucócitos/metabolismo , Proteínas Luminescentes/biossíntese , Pulmão/imunologia , Pulmão/metabolismo , Viabilidade Microbiana , Proteína Vermelha FluorescenteRESUMO
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.
