Hyperspectral Fluorescence Imaging with a New Polarity-Ultrasensitive Fluorescent Probe.

Area of Science:

• Chemical Biology
• Biomedical Imaging
• Molecular Probes

Background:

• Hyperspectral fluorescence imaging (HSFI) requires probes with high spectral sensitivity for simultaneous morphological and microenvironmental analysis.
• Current limitations in probe development hinder the full potential of HSFI.

Purpose of the Study:

• To develop a novel fluorescent probe with enhanced spectral sensitivity to microenvironmental polarity.
• To apply this probe for advanced HSFI in biological systems.
• To enable unprecedented visualization of lipid droplet dynamics.

Main Methods:

• Rational design of donor-π-acceptor molecules to optimize π-spacer effects on spectral sensitivity.
• Synthesis and characterization of the new fluorescent probe, Lipi-PS.
• Application of Lipi-PS in HSFI of cells, tissues, and zebrafishes using a 3D-SpecDIM system.

Main Results:

• Identified the critical role of the π-spacer in tuning wavelength sensitivity to polarity.
• Developed Lipi-PS, a probe with record-high wavelength sensitivity and lipid droplet targeting capabilities.
• Achieved 5D tracking (xyzλt) of individual lipid droplets, visualizing their polarity dynamics at the single-droplet level for the first time.

Conclusions:

• Lipi-PS significantly advances HSFI capabilities by providing superior spectral sensitivity.
• The developed probe and imaging method allow for detailed analysis of microenvironmental changes and lipid droplet behavior.
• This work opens new avenues for studying cellular processes and disease mechanisms involving lipid metabolism.