Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy
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 spe...
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| Format: | Article |
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MDPI AG
2022-02-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/3/1010 |
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| author | Roksana Markiewicz Jagoda Litowczenko Jacek Gapiński Anna Woźniak Stefan Jurga Adam Patkowski |
| author_facet | Roksana Markiewicz Jagoda Litowczenko Jacek Gapiński Anna Woźniak Stefan Jurga Adam Patkowski |
| author_sort | Roksana Markiewicz |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T23:27:13Z |
| format | Article |
| id | doaj.art-47e2d6e85a834059a99c024c63c3cc68 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T23:27:13Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-47e2d6e85a834059a99c024c63c3cc682023-11-23T17:16:09ZengMDPI AGMolecules1420-30492022-02-01273101010.3390/molecules27031010Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation SpectroscopyRoksana Markiewicz0Jagoda Litowczenko1Jacek Gapiński2Anna Woźniak3Stefan Jurga4Adam Patkowski5NanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, PolandNanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, PolandDepartment of Molecular Biophysics, Faculty of Physics, Adam Mickiewicz University in Poznan, ul. Uniwersytetu Poznańskiego 3, 61-614 Poznan, PolandNanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, PolandNanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, PolandNanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, PolandMeasurement 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.https://www.mdpi.com/1420-3049/27/3/1010nitric oxideNOnitric oxide determinationfluorescence correlation spectroscopyFCS |
| spellingShingle | Roksana Markiewicz Jagoda Litowczenko Jacek Gapiński Anna Woźniak Stefan Jurga Adam Patkowski Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy Molecules nitric oxide NO nitric oxide determination fluorescence correlation spectroscopy FCS |
| title | Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy |
| title_full | Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy |
| title_fullStr | Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy |
| title_full_unstemmed | Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy |
| title_short | Nanomolar Nitric Oxide Concentrations in Living Cells Measured by Means of Fluorescence Correlation Spectroscopy |
| title_sort | nanomolar nitric oxide concentrations in living cells measured by means of fluorescence correlation spectroscopy |
| topic | nitric oxide NO nitric oxide determination fluorescence correlation spectroscopy FCS |
| url | https://www.mdpi.com/1420-3049/27/3/1010 |
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