A measure of endosomal pH by flow cytometry in Dictyostelium.

BMC Research Notes
Anna MarchettiPierre Cosson

Abstract

Dictyostelium amoebae are frequently used to study the organization and function of the endocytic pathway, and specific protocols are essential to measure the dynamics of endocytic compartments and their internal pH. We have revisited these classical protocols to measure more accurately endosomal pH, making use of a fluorescent probe (Oregon green) more adequate for very acidic pH values. This pH-sensitive probe was combined with a pH-insensitive marker, in order to visualize simultaneously endosome dynamics and pH changes. Finally, a flow cytometer was used to measure endosomal pH in individual cells. Using these simple protocols the endosomal pH of endocytic compartments can be assessed accurately, revealing the extreme acidity of Dictyostelium lysosomes (pH <3.5).

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Citations

Sep 22, 2012·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Alireza ShalviriXiao Yu Wu
Jun 9, 2009·Journal of Biophotonics·Anja Mittag, Attila Tárnok
Nov 3, 2010·Cellular Microbiology·Emmanuelle LelongPierre Cosson
May 20, 2015·Magnetic Resonance in Medicine : Official Journal of the Society of Magnetic Resonance in Medicine·P Stephen PatrickKevin M Brindle
Apr 16, 2016·The Journal of Physical Chemistry. a·Magda A DallemagneNathan C Eddingsaas
Oct 7, 2014·Soft Matter·Fei LiuEugenia Kharlampieva
Jan 12, 2017·Cellular Microbiology·Jade LeibaPierre Cosson
Jul 4, 2017·Cells·Eunice Domínguez-MartínRicardo Escalante
Jul 3, 2013·Nature Communications·Yu-Hsien HungWei Yuan Yang
May 14, 2016·Journal of Cell Science·Aurélie GuehoFrançois Letourneur
Jan 22, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Sandrine DaubeufDenis Hudrisier
Feb 7, 2014·The Journal of Immunology : Official Journal of the American Association of Immunologists·Matthew WheelwrightPhilip T Liu
Jan 23, 2018·Frontiers in Immunology·Joe Dan DunnThierry Soldati
Jan 24, 2020·BMC Biology·Fu-Sheng ChangCatherine Pears
Jan 29, 2021·Frontiers in Cell and Developmental Biology·Johannes Westman, Sergio Grinstein
Feb 23, 2021·Frontiers in Cellular and Infection Microbiology·Cyril GuilhenPierre Cosson
Feb 24, 2012·Journal of Cell Science·Wanessa C LimaPierre Cosson
Mar 14, 2019·ACS Sensors·Sandra BurgstallerRoland Malli

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Methods Mentioned

BETA
Fluorescence
flow cytometry

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