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A pyrene-based two-photon excitable fluorescent probe to visualize nuclei in live cells
Chathura S Abeywickrama1, Kaveesha J Wijesinghe2, Caroline B Plescia3
1Department of Chemistry, University of Akron, 44325, Akron, Ohio, USA.
Summary
A novel pyrene-pyridinium dye visualizes live cell nuclei using two-photon fluorescence microscopy. This dye acts as a DNA intercalator, enabling efficient nucleus staining at low concentrations.
Area of Science:
- * Photochemistry and Photophysics
- * Molecular Imaging
- * Biophysical Chemistry
Background:
- * Two-photon spectroscopy offers advantages for biological imaging due to its deeper tissue penetration and reduced phototoxicity.
- * Developing novel fluorescent probes is crucial for advancing cellular imaging techniques.
- * Pyrene-based dyes are known for their photophysical properties, but their application in live-cell imaging requires further investigation.
Purpose of the Study:
- * To investigate the two-photon absorption properties of a novel pyrene-pyridinium dye.
- * To evaluate the dye's potential for cellular two-photon fluorescence microscopy imaging.
- * To elucidate the mechanism of nucleus staining and DNA interaction.
Main Methods:
- * Synthesis and characterization of the pyrene-pyridinium dye (probe 1).
- * Measurement of two-photon absorption cross-sections.
- * Live-cell imaging using two-photon fluorescence microscopy.
- * Spectroscopic studies to determine DNA interaction (intercalation).
Main Results:
- * Probe 1 exhibited significant two-photon absorption properties.
- * Effective visualization of live cell nuclei was achieved at a low probe concentration (1 μM).
- * Spectroscopic data indicated that probe 1 intercalates into DNA.
- * Nucleus staining efficiency correlated with substituents on the pyridinium fragment.
Conclusions:
- * The pyrene-pyridinium dye is a promising candidate for two-photon spectroscopy and live-cell nuclear imaging.
- * The dye's DNA intercalation mechanism underlies its nucleus-specific staining ability.
- * Further development of pyridinium substituents could optimize probe performance for specific imaging applications.
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