A Blue Light and Two-Photon Activatable Rhodamine Fluorophore

Area of Science:

• Chemical Biology
• Biophotonics
• Materials Science

Background:

• Photoactivatable fluorophores (PAFs) are crucial for spatiotemporal control in biological imaging.
• Existing PAFs often require UV irradiation, limiting their application range.
• There is a need for PAFs activatable by visible or near-infrared (NIR) light for deeper tissue penetration and reduced phototoxicity.

Purpose of the Study:

• To synthesize and characterize a novel rhodamine-based photoactivatable fluorophore.
• To investigate the photoreaction mechanism of the new fluorophore.
• To demonstrate the utility of the developed PAF in laser scanning microscopy for generating patterned illumination.

Main Methods:

• Synthesis of a novel rhodamine derivative.
• Photophysical characterization of the fluorophore's activation properties.
• Immobilization of the PAF within a hydrogel matrix.
• Demonstration using one-photon (1P) and two-photon (2P) laser scanning microscopy.

Main Results:

• Successful synthesis of a blue light (1P) and NIR light (2P) activatable rhodamine fluorophore.
• Detailed investigation of the underlying photoreaction mechanism.
• Demonstrated ability to write and read spatially resolved illumination patterns with high contrast in a hydrogel.
• Successful application in both 1P and 2P laser scanning microscopy.

Conclusions:

• The developed rhodamine-based PAF offers versatile activation options (blue and NIR light).
• This PAF allows for precise spatial control of fluorescence, enabling advanced imaging techniques.
• The hydrogel immobilization strategy facilitates robust pattern generation for microscopy applications.