Quantifying pH in Malaria Using pHluorin and Flow Cytometry

Jeffrey Agyapong; Petra Rohrbach

doi:10.1007/978-1-0716-3052-5_13

Quantifying pH in Malaria Using pHluorin and Flow Cytometry

Methods Mol Biol. 2023:2644:211-224. doi: 10.1007/978-1-0716-3052-5_13.

Authors

Jeffrey Agyapong¹, Petra Rohrbach²

Affiliations

¹ Institute of Parasitology, McGill University, Montreal, QC, Canada.
² Institute of Parasitology, McGill University, Montreal, QC, Canada. petra.rohrbach@mcgill.ca.

PMID: 37142924
DOI: 10.1007/978-1-0716-3052-5_13

Abstract

Intracellular pH (pH_i) plays a critical role in the regulation of numerous biological functions where specific pH ranges are required for optimal operation within cells. Slight pH changes can impact the regulation of diverse molecular processes, including enzymatic activities, ion channels, and transporters, which all play a role in cell functions. Methods for quantifying pH_i continue to evolve and include various optical methods using fluorescent pH indicators. Here, we provide a protocol to measure pH_i in the cytosol of Plasmodium falciparum blood stage parasites by means of flow cytometry and using pHluorin2, a pH-sensitive fluorescent protein that has been introduced into the genome of the parasite.

Keywords: Acidification; Alkalization; Flow cytometry; Fluorescence; Green fluorescent protein; Intracellular pH; Plasmodium; pH quantification; pHluorin2.

Publication types

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

MeSH terms

Coloring Agents
Flow Cytometry / methods
Green Fluorescent Proteins / genetics
Humans
Hydrogen-Ion Concentration
Malaria*
Plasmodium falciparum* / genetics
Plasmodium falciparum* / metabolism

Substances

PHluorin
Green Fluorescent Proteins
Coloring Agents