Fluorometric Na
Calibration
Cell Line, Tumor
Cytoplasm
/ metabolism
Flow Cytometry
/ methods
Fluorescence
Fluorescent Dyes
/ chemistry
Gramicidin
/ pharmacology
Humans
Ionophores
/ metabolism
Ions
Ouabain
/ pharmacology
Single-Cell Analysis
Sodium
/ metabolism
Sodium-Potassium-Exchanging ATPase
/ metabolism
Staurosporine
/ pharmacology
Flow cytometry; Na+-sensitive fluorescent probe; Cellular Na+ determination; Ionophores; Cellular Na+ flame emission assay
Journal
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
ISSN: 1421-9778
Titre abrégé: Cell Physiol Biochem
Pays: Germany
ID NLM: 9113221
Informations de publication
Date de publication:
29 May 2020
29 May 2020
Historique:
accepted:
18
05
2020
entrez:
29
5
2020
pubmed:
29
5
2020
medline:
13
1
2021
Statut:
ppublish
Résumé
Sodium is a key player in the fundamental cell functions. Fluorescent probes are indispensable tools for monitoring intracellular sodium levels in single living cells. Since the fluorescence of sodium-sensitive dyes in cells is significantly different from that in an aqueous solution, the fluorescence signal is calibrated in situ indirectly using ionophores for equalizing external and intracellular ion concentration. Attempts to compare data obtained using fluorescent probes and by direct flame emission analysis are sparse and results are inaccurate. We determined the intracellular sodium concentration in U937 cells by flow cytometry using the Na It is revealed that both methods are comparable when intracellular sodium concentration was modified by ouabain-mediated blockage of the sodium pump or staurosporine-induced apoptosis. The ANG fluorescence of cells treated with ionophores is approximately two times lower than that in cells with the same Na The sodium sensitive dye ANG-2 is a sensitive and useful probe for determination changes in Na
Sections du résumé
BACKGROUND/AIMS
OBJECTIVE
Sodium is a key player in the fundamental cell functions. Fluorescent probes are indispensable tools for monitoring intracellular sodium levels in single living cells. Since the fluorescence of sodium-sensitive dyes in cells is significantly different from that in an aqueous solution, the fluorescence signal is calibrated in situ indirectly using ionophores for equalizing external and intracellular ion concentration. Attempts to compare data obtained using fluorescent probes and by direct flame emission analysis are sparse and results are inaccurate.
METHODS
METHODS
We determined the intracellular sodium concentration in U937 cells by flow cytometry using the Na
RESULTS
RESULTS
It is revealed that both methods are comparable when intracellular sodium concentration was modified by ouabain-mediated blockage of the sodium pump or staurosporine-induced apoptosis. The ANG fluorescence of cells treated with ionophores is approximately two times lower than that in cells with the same Na
CONCLUSION
CONCLUSIONS
The sodium sensitive dye ANG-2 is a sensitive and useful probe for determination changes in Na
Substances chimiques
Fluorescent Dyes
0
Ionophores
0
Ions
0
Gramicidin
1405-97-6
Ouabain
5ACL011P69
Sodium
9NEZ333N27
Sodium-Potassium-Exchanging ATPase
EC 7.2.2.13
Staurosporine
H88EPA0A3N
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
556-566Subventions
Organisme : State assignment of Russian Federation
ID : 0124-2019-0003
Pays : Russian Federation
Organisme : Institute of Cytology of RAS
Pays : Russian Federation
Informations de copyright
© Copyright by the Author(s). Published by Cell Physiol Biochem Press.
Déclaration de conflit d'intérêts
The authors declare no competing interests.