DCDHF fluorophores for single-molecule imaging in cells

Area of Science:

• Chemical Biology
• Microscopy
• Organic Chemistry

Background:

• Small-molecule fluorescent labels are crucial for live-cell single-molecule imaging.
• Existing labels often fail to meet stringent requirements like photostability, solubility, and specific spectral properties.

Purpose of the Study:

• To design and characterize novel DCDHF (2-dicyanomethylene-3-cyano-2,5-dihydrofuran) fluorophores for enhanced live-cell imaging.
• To develop photoactivatable variants for super-resolution microscopy applications.

Main Methods:

• Utilized a push-pull molecular design with amine donor and DCDHF acceptor groups linked by a conjugated system.
• Synthesized and characterized DCDHF fluorophores, evaluating their photophysical properties, solubility, and membrane permeability.
• Engineered a red-emitting DCDHF fluorophore for photoactivation capabilities.

Main Results:

• Developed a series of DCDHF fluorophores with tunable absorption, emission, solubility, and membrane affinity.
• Demonstrated high photostability and environmental sensitivity of the DCDHF fluorophores.
• Created a novel photoactivatable DCDHF fluorophore that is initially dark and can be activated by violet light.

Conclusions:

• DCDHF fluorophores represent a versatile new class of probes for live-cell imaging.
• The photoactivatable DCDHF fluorophore enables precise control over single-molecule emission for super-resolution techniques.
• These fluorophores address key limitations in current single-molecule imaging technologies.