Fluorescence Lifetime Measurements and Analyses: Protocols Using Flow Cytometry and High-Throughput Microscopy

Area of Science:

• Biophysics
• Cell Biology
• Analytical Chemistry

Background:

• Fluorescence decay timing is a well-established parameter in cell analysis.
• Traditional cytometry methods benefit from incorporating fluorescence lifetime measurements.
• Understanding fluorescence decay kinetics is crucial for advanced cytometric applications.

Purpose of the Study:

• To provide an overview of fluorescence lifetime measurement techniques in cytometry.
• To explore how fluorescence lifetime enhances existing cytometry protocols.
• To review modern high-throughput lifetime analysis methods.

Main Methods:

• Review of theoretical background and history of time-resolved flow cytometry.
• Analysis of applications benefiting from fluorescence lifetime as a photophysical trait.
• Discussion of modern high-throughput scanning techniques including high-speed microscopy and machine learning.

Main Results:

• Fluorescence lifetime provides quantitative, informative data for cell analysis.
• Time-resolved flow cytometry has evolved significantly for enhanced cellular insights.
• Modern techniques enable high-throughput fluorescence lifetime scanning.

Conclusions:

• Fluorescence lifetime measurement is a valuable cytometric parameter.
• Advanced techniques like machine learning are revolutionizing lifetime analysis.
• This approach offers significant potential for future cell biology research.