Time‐Gated FRET Nanoprobes for Autofluorescence‐Free Long‐Term In Vivo Imaging of Developing Zebrafish

• DOI: 10.1002/adma.202003912
• Publication Date: 2020-08-28T12:56:49Z
• Authors: Marcelina Cardoso...; Ingrid Colin; Gabriel Ribeiro D...; Kimihiro Susumu; Michaël Demarque; Igor L. Medintz; Niko Hildebrandt
• Keywords: Autofluorescence; Photobleaching; Nanoprobe; Fluorescence-lifetime imaging microscopy; Live cell imaging

Abstract The zebrafish is an important vertebrate model for disease, drug discovery, toxicity, embryogenesis, and neuroscience. In vivo fluorescence microscopy can reveal cellular subcellular details down to the molecular level with fluorescent proteins (FPs) currently main tool imaging. However, long maturation times, low brightness, photobleaching, broad emission spectra, sample autofluorescence are disadvantages that cannot be easily overcome by FPs. Here, a bright photostable terbium‐to‐quantum dot (QD) Förster resonance energy transfer (FRET) nanoprobe narrow tunable bands intracellular in imaging presented. photoluminescence (PL) lifetime enables time‐gated (TG) detection without background. Intracellular four‐color multiplexing single excitation wavelength situ assembly FRET mCherry demonstrate versatility of TG‐FRET nanoprobes possibility bioconjugation FPs combined nanoprobe‐FP sensing. Upon injection at one‐cell stage, imaged developing embryos over seven days toxicity similar injected RNA strongly improved signal‐to‐background ratios compared non‐TG This work provides strategy advancing applications beyond capabilities