Tempo-spectral multiplexing in flow cytometry with lifetime detection using QD-encoded polymer beads

Area of Science:

• Materials Science
• Biotechnology
• Analytical Chemistry

Background:

• Semiconductor quantum dots (QDs) in polymer microbeads are promising for spectral multiplexing and color encoding.
• Luminescence lifetimes of QDs, rarely used for encoding, offer unique time ranges inaccessible to other luminophores.

Purpose of the Study:

• To demonstrate the potential of II/VI semiconductor QDs for expanding lifetime encoding in multiplexing applications.
• To integrate QD-based lifetime encoding with existing methods for enhanced multiplexing capabilities.

Main Methods:

• Preparation and characterization of two types of QD-loaded microbeads using photoluminescence and single-particle confocal laser scanning microscopy.
• Integration of QD-encoded beads with dye-encoded microparticles for multiplexing studies.
• Utilizing a novel time-domain luminescence lifetime flow cytometer (LT-FCM setup).

Main Results:

• QD-loaded microbeads exhibited luminescence lifetimes in the several 10s of nanoseconds range.
• Successful combination of QD-encoded and dye-encoded particles demonstrated increased lifetime codes.
• Demonstrated potential for tempo-spectral multiplexing by combining time and color domains.

Conclusions:

• II/VI semiconductor QDs significantly expand the range of luminescence lifetimes available for encoding in multiplexing.
• The developed LT-FCM approach enables enhanced multiplexing by combining spectral and temporal information.
• This technology offers a powerful alternative for advanced multiplexing in flow cytometry and related fields.