ArticleOptics express2024
Multiplexed imaging in live cells using pulsed interleaved excitation spectral FLIM.
Article in Optics express, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- A Chemigenetic Fluorescence Lifetime Biosensor for Dopamine Sensing.bioRxiv : the preprint server for biology · 2026Article
- Phasor-Based Spatio-Spectral Segmentation for Hyperspectral Fluorescence Microscopy.Journal of biophotonics · 2026Article
- Lifetime-based multiplexed detection of viral RNA using fluorogenic aptamers.bioRxiv : the preprint server for biology · 2026Article
- Vessel-on-a-Chip to Study Vascular Endothelial Inflammation.Current protocols · 2025Article
- Two-photon autofluorescence lifetime assay of rabbit photoreceptors and retinal pigment epithelium during light-dark visual cycles in rabbit retina.Biomedical optics express · 2024Article
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Authors and funding
11 authors.
Funding
Abstract
Multiplexed fluorescence detection has become increasingly important in the fields of biosensing and bioimaging. Although a variety of excitation/detection optical designs and fluorescence unmixing schemes have been proposed to allow for multiplexed imaging, rapid and reliable differentiation and quantification of multiple fluorescent species at each imaging pixel is still challenging. Here we present a pulsed interleaved excitation spectral fluorescence lifetime microscopic (PIE-sFLIM) system that can simultaneously image six fluorescent tags in live cells in a single hyperspectral snapshot. Using an alternating pulsed laser excitation scheme at two different wavelengths and a synchronized 16-channel time-resolved spectral detector, our PIE-sFLIM system can effectively excite multiple fluorophores and collect their emission over a broad spectrum for analysis. Combining our system with the advanced live-cell labeling techniques and the lifetime/spectral phasor analysis, our PIE-sFLIM approach can well unmix the fluorescence of six fluorophores acquired in a single measurement, thus improving the imaging speed in live-specimen investigation.
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Registered trials
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