Resolving subcellular pH with a quantitative fluorescent lifetime biosensor.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
12 10 2022
12 10 2022
Historique:
received:
23
11
2021
accepted:
13
09
2022
entrez:
12
10
2022
pubmed:
13
10
2022
medline:
15
10
2022
Statut:
epublish
Résumé
Changes in sub-cellular pH play a key role in metabolism, membrane transport, and triggering cargo release from therapeutic delivery systems. Most methods to measure pH rely on intensity changes of pH sensitive fluorophores, however, these measurements are hampered by high uncertainty in the inferred pH and the need for multiple fluorophores. To address this, here we combine pH dependant fluorescent lifetime imaging microscopy (pHLIM) with deep learning to accurately quantify sub-cellular pH in individual vesicles. We engineer the pH sensitive protein mApple to localise in the cytosol, endosomes, and lysosomes, and demonstrate that pHLIM can rapidly detect pH changes induced by drugs such as bafilomycin A1 and chloroquine. We also demonstrate that polyethylenimine (a common transfection reagent) does not exhibit a proton sponge effect and had no measurable impact on the pH of endocytic vesicles. pHLIM is a simple and quantitative method that will help to understand drug action and disease progression.
Identifiants
pubmed: 36224168
doi: 10.1038/s41467-022-33348-z
pii: 10.1038/s41467-022-33348-z
pmc: PMC9556823
doi:
Substances chimiques
Protons
0
Chloroquine
886U3H6UFF
Polyethyleneimine
9002-98-6
Banques de données
figshare
['10.6084/m9.figshare.20454867.v1']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6023Informations de copyright
© 2022. The Author(s).
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