Photoswitchable polyynes for multiplexed stimulated Raman scattering microscopy with reversible light control

Area of Science:

• Biophysics
• Chemical Biology
• Optical Microscopy

Background:

• Optical imaging is crucial for biological research, with stimulated Raman scattering (SRS) microscopy offering high chemical specificity.
• Existing SRS methods lack multiplexing capabilities with reversible photocontrol for dynamic cellular studies.

Purpose of the Study:

• To develop a photoswitchable probe for multiplexed SRS microscopy with reversible photocontrol.
• To demonstrate advanced imaging of cellular dynamics in living systems.

Main Methods:

• Creation of Carbow-switch, a photoswitchable polyyne probe by coupling diarylethene with super-multiplexed Carbow.
• Optimization of electronic and vibrational spectroscopy for enhanced photoswitching and SRS response.
• Application in reversible, spatial-selective multiplexed SRS imaging of organelles in living cells.

Main Results:

• Carbow-switch demonstrated excellent photoswitching under visible light with significant SRS signal enhancement.
• Achieved reversible and spatial-selective multiplexed SRS imaging of various organelles in living cells.
• Enabled photo-selective time-lapse imaging of organelle dynamics during oxidative stress and protein phase separation.

Conclusions:

• Carbow-switch provides a powerful tool for photoswitchable SRS microscopy.
• This technology enables new possibilities for studying complex cellular interactions and dynamics with high spatiotemporal precision.