On-the-fly decoding luminescence lifetimes in the microsecond region for lanthanide-encoded suspension arrays

Abstract

Significant multiplexing capacity of optical time-domain coding has been recently demonstrated by tuning luminescence lifetimes of the upconversion nanoparticles called ‘τ-Dots’. It provides a large dynamic range of lifetimes from microseconds to milliseconds, which allows creating large libraries of nanotags/microcarriers. However, a robust approach is required to rapidly and accurately measure the luminescence lifetimes from the relatively slow-decaying signals. Here we show a fast algorithm suitable for the microsecond region with precision closely approaching the theoretical limit and compatible with the rapid scanning cytometry technique. We exploit this approach to further extend optical time-domain multiplexing to the downconversion luminescence, using luminescence microspheres wherein lifetimes are tuned through luminescence resonance energy transfer. We demonstrate real-time discrimination of these microspheres in the rapid scanning cytometry, and apply them to the multiplexed probing of pathogen DNA strands. Our results indicate that tunable luminescence lifetimes have considerable potential in high-throughput analytical sciences.

White-light upconversion emission of lanthanide double-doped oxide nanoparticles via defect state luminescence of ZnO

“…However, the relatively low efficiency of white UCL sources still poses a challenge to researchers today. It is well-known that the efficiency of white UCL is highly dependent on host matrix, particle morphology, crystallite phase, and lanthanide dopants [18][19][20][21][22][23]. Therefore, the study of their host matrix is important for obtaining a good white UCL emission and white light [24].…”

High-Precision Pinpointing of Luminescent Targets in Encoder-Assisted Scanning Microscopy Allowing High-Speed Quantitative Analysis

“…We have also shown recently that luminescence lifetimes of lanthanide-based upconversion materials can be fine-tuned across the microsecond to millisecond range, allowing for creation of temporally multiplexed codes for luminescence detaction 33,37 . In parallel we have developed a controlled synthesis approach for bottom-up production of a library of colour-barcoded heterogeneous micro-rods at low cost 38 .…”

“…One solution to this problem includes the use of lanthanide luminescent materials exhibiting long lifetimes and/or photon upconversion properties, which are highly useful as either high-contrast molecular probes for direct labelling [24][25][26][27][28][29][30] or microsphere-based suspension arrays for high throughput assays [31][32][33] .…”

Phasor plotting with frequency-domain flow cytometry

“…We and others have demonstrated different versions of time-resolved flow cytometry [3,[7][8][9][10][11][12][13][14], and the main objective for augmenting flow cytometers to detect fluorescence lifetimes is to enhance cytometric data with a quantitative parameter that is independent of the measured fluorescence intensity [11,[15][16][17]. As a parameter, fluorescence lifetime can be used to discriminate among spectrally overlapping fluorescence signals as well as to validate the fluorescence intensity changes that arise from quenched fluorophores such as during Förster resonance energy transfer (FRET) events like those arising from changes in association state of tagged cytoplasmic proteins.…”