In Vivo Monitoring of Cytosolic pH Using the Ratiometric pH Sensor pHluorin

Tânia R Fernandes; Antonio Serrano; Antonio Di Pietro

doi:10.1007/978-1-0716-1795-3_9

In Vivo Monitoring of Cytosolic pH Using the Ratiometric pH Sensor pHluorin

Methods Mol Biol. 2022:2391:99-107. doi: 10.1007/978-1-0716-1795-3_9.

Authors

Tânia R Fernandes^{1

2}, Antonio Serrano^{1

3}, Antonio Di Pietro⁴

Affiliations

¹ Departamento de Genética, Campus de Excelencia Internacional Agroalimentario ceiA3, Universidad de Córdoba, Córdoba, Spain.
² GreenUPorto, Sustainable Agrifood Production Research Centre / Inov4Agro & DGAOT, Faculty of Sciences, University of Porto, Porto, Portugal.
³ Université Côte D'Azur, CNRS, INSERM, Institute of Biology Valrose (iBV), Nice, France.
⁴ Departamento de Genética, Campus de Excelencia Internacional Agroalimentario ceiA3, Universidad de Córdoba, Córdoba, Spain. ge2dipia@uco.es.

PMID: 34686980
DOI: 10.1007/978-1-0716-1795-3_9

Abstract

Cytosolic pH (pH_cyt) is a key factor controlling cell fate. The genetically encoded pH-sensor pHluorin has proven highly valuable for studies on pH_cyt in many living organisms. pHluorin displays a bimodal excitation spectrum with peaks at 395 nm and 475 nm, which is dependent on pH. Here we describe two different protocols for determining pH_cyt in the soil-borne fungal pathogen Fusarium oxysporum, based either on population or single-cell analysis.

Keywords: Confocal microscope; Cytosolic pH; Ratiometric pH sensor; Spectrofluorometer; pHluorin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytosol*
Fusarium
Green Fluorescent Proteins
Hydrogen-Ion Concentration

Substances

PHluorin
Green Fluorescent Proteins

Supplementary concepts

Fusarium oxysporum