Hyperdimensional Imaging Contrast Using an Optical Fiber.

Area of Science:

• Biophysics
• Microscopy
• Molecular Imaging

Background:

• Molecular fluorescence properties are crucial for studying biological processes.
• Hyperdimensional imaging microscopy (HDIM) integrates physical properties and biochemical states of fluorophores.
• Existing techniques may lack sufficient contrast for complex biological samples.

Purpose of the Study:

• To present a novel fiber-based imaging system for generating hyper-dimensional contrast.
• To combine multiple fluorescence properties into a single fluorescence lifetime decay curve.
• To demonstrate enhanced contrast for improved molecular differentiation.

Main Methods:

• Development of a fiber-based imaging system integrating controlled excitation polarization and temporally dispersed emission.
• Utilizing fluorescence lifetime imaging microscopy (FLIM) to generate spectrally coded, polarization-filtered lifetime distributions.
• Applying multiphoton excitation for imaging *Arabidopsis thaliana* epidermal cells.

Main Results:

• The system generates hyper-dimensional contrast by combining multiple fluorescence properties.
• Achieved superior contrast between different molecules compared to individual techniques.
• Demonstrated higher contrast in *Arabidopsis thaliana* epidermal cells using intrinsic anthocyanin emission.

Conclusions:

• The developed HDIM scheme offers better molecular contrast than traditional methods.
• The fiber-based system is simpler, modular, cost-efficient, and adaptable to existing FLIM microscopes.
• Contrast-based imaging is valuable for investigating cellular heterogeneity in biological samples.