Readily Accessible and Predictable Naphthalene-Based Two-Photon Fluorophore with Full Visible-Color Coverage

Area of Science:

• Organic Chemistry
• Biomedical Imaging
• Fluorescence Spectroscopy

Background:

• Developing novel fluorophores is crucial for advancing biological imaging techniques.
• Two-photon microscopy offers advantages in deep tissue imaging due to reduced phototoxicity and enhanced spatial resolution.
• Existing fluorophores may have limitations in spectral coverage, brightness, or specific sensing capabilities.

Purpose of the Study:

• To synthesize and characterize a new series of acedan-derived two-photon fluorophores.
• To achieve full coverage of the visible emission spectrum with these novel fluorophores.
• To demonstrate the application of these fluorophores in multicolor imaging and as a sensor for endogenous hydrogen peroxide (H2O2).

Main Methods:

• Computational prediction of emission wavelengths.
• Chemical synthesis of 22 acedan-derived fluorophores.
• Two-photon excitation microscopy for multicolor imaging.
• Development of a turn-on fluorescence sensor for H2O2 detection.

Main Results:

• Successfully synthesized 22 acedan-derived fluorophores with high synthetic feasibility.
• Achieved full coverage of visible wavelength emission, enabling multicolor imaging with single-wavelength excitation.
• Demonstrated successful visualization of multicolor images in biological samples.
• Designed and validated a turn-on two-photon fluorescence sensor for endogenous H2O2 in Raw 264.7 macrophage cells and rat brain hippocampus ex vivo.

Conclusions:

• The developed acedan-derived fluorophores offer a versatile platform for advanced biological imaging.
• Computational prediction proved effective in guiding the design of fluorophores with desired spectral properties.
• The novel sensor provides a valuable tool for studying H2O2 dynamics in biological systems.