Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy.
FCS
NO
fluorescence correlation spectroscopy
nitric oxide
nitric oxide determination
Journal
Molecules (Basel, Switzerland)
ISSN: 1420-3049
Titre abrégé: Molecules
Pays: Switzerland
ID NLM: 100964009
Informations de publication
Date de publication:
02 Feb 2022
02 Feb 2022
Historique:
received:
12
01
2022
revised:
28
01
2022
accepted:
31
01
2022
entrez:
15
2
2022
pubmed:
16
2
2022
medline:
23
2
2022
Statut:
epublish
Résumé
Measurement of the nitric oxide (NO) concentration in living cells in the physiological nanomolar range is crucial in understanding NO biochemical functions, as well as in characterizing the efficiency and kinetics of NO delivery by NO-releasing drugs. Here, we show that fluorescence correlation spectroscopy (FCS) is perfectly suited for these purposes, due to its sensitivity, selectivity, and spatial resolution. Using the fluorescent indicators, diaminofluoresceins (DAFs), and FCS, we measured the NO concentrations in NO-producing living human primary endothelial cells, as well as NO delivery kinetics, by an external NO donor to the immortal human epithelial living cells. Due to the high spatial resolution of FCS, the NO concentration in different parts of the cells were also measured. The detection of nitric oxide by means of diaminofluoresceins is much more efficient and faster in living cells than in PBS solutions, even though the conversion to the fluorescent form is a multi-step reaction.
Identifiants
pubmed: 35164275
pii: molecules27031010
doi: 10.3390/molecules27031010
pmc: PMC8838662
pii:
doi:
Substances chimiques
Fluorescent Dyes
0
Nitric Oxide
31C4KY9ESH
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Science Center
ID : 2014/15/B/ST5/04259
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