Multimodal Raman and Fluorescence Lifetime Imaging: A New Analytical Approach for Resolving Timing-Dependent...

Area of Science:

• Biomedical Optics
• Chemical Biology
• Vascular Biology

Background:

• Endothelial dysfunction disrupts vascular homeostasis via metabolic disturbances.
• Temporal recovery patterns of endothelial dysfunction are analytically underexplored.
• Objective evaluation of therapeutic efficacy requires precise molecular profiling.

Purpose of the Study:

• To develop and apply a high-content analytical platform for quantitative molecular mapping of endothelial dysfunction.
• To identify spectral biomarkers and metabolic phenotypes associated with endothelial dysfunction.
• To investigate the timing-dependent efficacy of enalaprilat in restoring endothelial function.

Main Methods:

• Integration of spontaneous Raman scattering, stimulated Raman scattering, coherent anti-Stokes Raman scattering, and fluorescence lifetime imaging microscopy.
• Multimodal imaging for simultaneous assessment of lipid metabolism, biochemical composition, and cellular bioenergetics.
• Chemometric analysis including principal component analysis and partial least-squares regression for spectral data interpretation.

Main Results:

• Identification of distinct spectral biomarkers (Raman bands at 701, 1748, 2884, 3011 cm⁻¹) and nonspectral descriptors (reduced cell volume, accelerated lipid droplet formation).
• Demonstration of a pronounced timing-dependent response to enalaprilat treatment.
• Preventive enalaprilat treatment preserved metabolic integrity, while post-treatment resulted in partial restitution and divergent metabolic trajectories.

Conclusions:

• Early intervention in endothelial dysfunction leads to analytically distinguishable molecular states.
• Multimodal spectroscopic and chemometric framework resolves subtle metabolic phenotypes.
• Temporal profiling is crucial for refining strategies in endothelial diagnostics and evaluating therapeutic efficacy.