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Article

Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy

by
Roksana Markiewicz
1,*,
Jagoda Litowczenko
1,*,
Jacek Gapiński
2,
Anna Woźniak
1,3,
Stefan Jurga
1 and
Adam Patkowski
1,2
1
NanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, Poland
2
Department of Molecular Biophysics, Faculty of Physics, Adam Mickiewicz University in Poznan, ul. Uniwersytetu Poznańskiego 3, 61-614 Poznan, Poland
3
Department of Biochemistry and Biotechnology, Poznan University of Life Sciences, Dojazd 11, 60-632 Poznan, Poland
*
Authors to whom correspondence should be addressed.
Molecules 2022, 27(3), 1010; https://doi.org/10.3390/molecules27031010
Submission received: 12 January 2022 / Revised: 28 January 2022 / Accepted: 31 January 2022 / Published: 2 February 2022
(This article belongs to the Special Issue NanoTech Poland 2021—Nanotechnology in Energy, Electronics, Photonics, Environment and Biomedicine)
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Abstract

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.
Keywords: nitric oxide; NO; nitric oxide determination; fluorescence correlation spectroscopy; FCS nitric oxide; NO; nitric oxide determination; fluorescence correlation spectroscopy; FCS

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MDPI and ACS Style

Markiewicz, R.; Litowczenko, J.; Gapiński, J.; Woźniak, A.; Jurga, S.; Patkowski, A. Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy. Molecules 2022, 27, 1010. https://doi.org/10.3390/molecules27031010

AMA Style

Markiewicz R, Litowczenko J, Gapiński J, Woźniak A, Jurga S, Patkowski A. Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy. Molecules. 2022; 27(3):1010. https://doi.org/10.3390/molecules27031010

Chicago/Turabian Style

Markiewicz, Roksana, Jagoda Litowczenko, Jacek Gapiński, Anna Woźniak, Stefan Jurga, and Adam Patkowski. 2022. "Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy" Molecules 27, no. 3: 1010. https://doi.org/10.3390/molecules27031010

APA Style

Markiewicz, R., Litowczenko, J., Gapiński, J., Woźniak, A., Jurga, S., & Patkowski, A. (2022). Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy. Molecules, 27(3), 1010. https://doi.org/10.3390/molecules27031010

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