9-Cyanopyronin probe palette for super-multiplexed vibrational imaging

Area of Science:

• Chemical Biology
• Optical Imaging
• Spectroscopy

Background:

• Multiplexed optical imaging is crucial for visualizing complex biological interactions.
• Vibrational microscopy offers high specificity due to sharp spectral linewidths.
• Previous work introduced Manhattan Raman Scattering (MARS) probes coupled with epr-SRS microscopy.

Purpose of the Study:

• To develop robust synthetic methods for creating diverse MARS probes.
• To establish a predictive model for rational dye design based on structural-vibrational frequency correlations.
• To expand the utility of MARS probes for advanced multiplexed vibrational imaging.

Main Methods:

• Developed versatile synthetic routes for MARS probes with varied core atoms, ring structures, and isotopic substitutions.
• Discovered a predictive model linking MARS probe structure to vibrational frequencies.
• Integrated the expanded MARS probe library with electronic pre-resonant stimulated Raman scattering (epr-SRS) microscopy.

Main Results:

• Successfully synthesized a diverse library of MARS probes with tunable Raman shifts.
• Demonstrated the predictive model's efficacy in guiding the rational design of new MARS dyes.
• Showcased versatile labeling modalities and significantly increased multiplexing capacity using MARS probes with epr-SRS microscopy.

Conclusions:

• The developed synthetic methodology and predictive model enable the rational design of novel MARS probes.
• The expanded MARS probe library enhances the capabilities of epr-SRS microscopy for multiplexed imaging.
• This work paves the way for next-generation vibrational imaging with broader applications in biological research.