Lifetime-encoded microparticles - application to flow cytometry
- Analytical methods require efficient and versatile strategies to measure an increasing number of analytes that can be used in conjunction with established platforms like flow cytometry. Spectral multiplexing suffers from problems such as spectral crosstalk and often requires different excitation light sources increasing instrumentation costs. Thus, the number of distinguishable reporters with intensity-based barcodes is limited. An alternative can be lifetime encoding for discrimination of fluorophores based on their fluorescence decay kinetics.
We report on the suitability of µm-sized polymer particles stained with organic dyes for lifetime encoding. These dyes are excitable at a standard laser diode wavelength and detectable within a single spectral window. For lifetime-based discrimination, these dyes display sufficiently different luminescence decay kinetics. We present the spectroscopic properties of these beads and address challenges like the limited number of detectableAnalytical methods require efficient and versatile strategies to measure an increasing number of analytes that can be used in conjunction with established platforms like flow cytometry. Spectral multiplexing suffers from problems such as spectral crosstalk and often requires different excitation light sources increasing instrumentation costs. Thus, the number of distinguishable reporters with intensity-based barcodes is limited. An alternative can be lifetime encoding for discrimination of fluorophores based on their fluorescence decay kinetics.
We report on the suitability of µm-sized polymer particles stained with organic dyes for lifetime encoding. These dyes are excitable at a standard laser diode wavelength and detectable within a single spectral window. For lifetime-based discrimination, these dyes display sufficiently different luminescence decay kinetics. We present the spectroscopic properties of these beads and address challenges like the limited number of detectable photons in a flow for the reliable discrimination. These studies are expected to pave the road to new applications of fluorescence lifetime multiplexing for time-domain flow cytometry.…