High-speed multiparameter photophysical analyses of fluorophore libraries

Area of Science:

• Biophysics
• Microfluidics
• Optical Engineering

Background:

• High-throughput screening of fluorescent probes is essential for advancing imaging and biosensor technologies.
• Existing methods for photophysical characterization are often slow and cannot handle large probe libraries.

Purpose of the Study:

• To develop a rapid, high-throughput method for multiparameter photophysical measurements of fluorescent probe libraries.
• To enable simultaneous assessment of fluorescence lifetime and photobleaching for imaging performance prediction.

Main Methods:

• A novel microfluidic flow cytometer was designed and implemented.
• The system assays large cell-based fluorophore libraries (10^4-10^5 members).
• Simultaneous measurement of fluorescence lifetime and photobleaching rates.

Main Results:

• Demonstrated rapid assaying of large-scale fluorophore libraries.
• Successfully resolved diverse photophysical characteristics, including fluorescence lifetime and photobleaching.
• Identified rare populations within the libraries based on photophysical properties.

Conclusions:

• The microfluidic flow cytometer provides a powerful tool for high-speed multiparameter photophysical analysis.
• This technology significantly accelerates the development and optimization of fluorescent probes for imaging and biosensors.
• Enables efficient screening and selection of optimal fluorophores from large libraries.