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bioRxiv · 10.1101/2025.08.13.670148

Single-dye, transfection-free FLIM multiplexing via bioorthogonal chemistry

Dadina, N.·Kwon, J. H.·Lesiak, L.·Zheng, S.·Zoltek, M.·Brauer, D.·Schepartz, A.
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Abstract

Fluorescence lifetime imaging microscopy (FLIM) can visualize multiple targets in a single spectral window, making it a powerful tool to overcome multiplexing limitations during live cell fluorescence microscopy. Here we show that small molecule probes-which are well-suited for imaging applications due to high specificity, low toxicity, and the elimination of transfection requirements-can be fine-tuned via bioorthogonal chemistry to exhibit predictably different fluorescent lifetimes suitable for FLIM multiplexing.

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Dadina, N., Kwon, J. H., Lesiak, L., Zheng, S., Zoltek, M., Brauer, D., Schepartz, A.. 2025-08-16. Single-dye, transfection-free FLIM multiplexing via bioorthogonal chemistry. https://doi.org/10.1101/2025.08.13.670148

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