title: Biocompatible photoresistant far-red emitting, fluorescent polymer probes, with near-infrared two-photon absorption, for living cell and zebrafish embryo imaging.
abstract: Exogenous probes with far-red or near-infrared (NIR) two-photon absorption and fluorescence emission are highly desirable for deep tissue imaging while limiting autofluorescence. However, molecular probes exhibiting such properties are often hydrophobic. As an attractive alternative, we synthesized water-soluble polymer probes carrying multiple far-red fluorophores and demonstrated here their potential for live cell and zebrafish embryo imaging. First, at concentrations up to 10 μm, these polymer probes were not cytotoxic. They could efficiently label living HeLa cells, T lymphocytes and neurons at an optimal concentration of 0.5 μm. Moreover, they exhibited a high resistance to photobleaching in usual microscopy conditions. In addition, these polymer probes could be successfully used for in toto labeling and in vivo two-photon microscopy imaging of developing zebrafish embryos, with remarkable properties in terms of biocompatibility, internalization, diffusion, stability and wavelength emission range. The near-infrared two-photon absorption peak at 910 nm is particularly interesting since it does not excite the zebrafish endogenous fluorescence and is likely to enable long-term time-lapse imaging with limited photodamage.
authors: École Normale Supérieure de Lyon, Laboratoire Joliot-Curie, CNRS USR3010, F-69364 Lyon, France; INSA-Lyon, Laboratoire Ingénierie des Matériaux Polymères, CNRS UMR5223, Université Claude Bernard Lyon 1, F-69621 Villeurbanne, France.
authors: Salim
authors: S
authors: Adjili
authors: École Normale Supérieure de Lyon, Laboratoire Joliot-Curie, CNRS USR3010, F-69364 Lyon, France; INSA-Lyon, Laboratoire Ingénierie des Matériaux Polymères, CNRS UMR5223, Université Claude Bernard Lyon 1, F-69621 Villeurbanne, France. Electronic address: arnaud.favier@ens-lyon.fr.
authors: Arnaud
authors: A
authors: Favier
authors: École Normale Supérieure de Lyon, Unité de Virologie Humaine, INSERM U758, Université Claude Bernard Lyon 1, F-69364 Lyon, France.
authors: Guillaume
authors: G
authors: Fargier
authors: École Normale Supérieure de Lyon, Unité de Virologie Humaine, INSERM U758, Université Claude Bernard Lyon 1, F-69364 Lyon, France.
authors: Audrey
authors: A
authors: Thomas
authors: École Normale Supérieure de Lyon, Laboratoire de Chimie, CNRS UMR5182, Université Claude Bernard Lyon 1, Site Monod, 46 allée d'Italie, F-69364 Lyon, France.
authors: Julien
authors: J
authors: Massin
authors: École Normale Supérieure de Lyon, Laboratoire Joliot-Curie, CNRS USR3010, F-69364 Lyon, France.
authors: Karine
authors: K
authors: Monier
authors: Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé, CNRS UMR5236, F-34293 Montpellier, France.
authors: Cyril
authors: C
authors: Favard
authors: SFR Santé Lyon Est-CIQLE, CNRS UMS3453/INSERM US7, Université Claude Bernard Lyon1, F-69372 Lyon, France.
authors: Christophe
authors: C
authors: Vanbelle
authors: UMS AMAGEN, CNRS UMS3504/INRA UMS1374, Bat 32, Avenue de la Terrasse, F-91198 Gif-sur-Yvette, France; BioEmergences, CNRS UPS3674, Bat 32, Avenue de la Terrasse, F-91198 Gif-sur-Yvette, France.
authors: Sylvia
authors: S
authors: Bruneau
authors: BioEmergences, CNRS UPS3674, Bat 32, Avenue de la Terrasse, F-91198 Gif-sur-Yvette, France.
authors: Nadine
authors: N
authors: Peyriéras
authors: École Normale Supérieure de Lyon, Laboratoire de Chimie, CNRS UMR5182, Université Claude Bernard Lyon 1, Site Monod, 46 allée d'Italie, F-69364 Lyon, France.
authors: Chantal
authors: C
authors: Andraud
authors: École Normale Supérieure de Lyon, Unité de Virologie Humaine, INSERM U758, Université Claude Bernard Lyon 1, F-69364 Lyon, France; Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé, CNRS UMR5236, F-34293 Montpellier, France. Electronic address: delphine.muriaux@cpbs.cnrs.fr.
authors: Delphine
authors: D
authors: Muriaux
authors: École Normale Supérieure de Lyon, Laboratoire Joliot-Curie, CNRS USR3010, F-69364 Lyon, France; INSA-Lyon, Laboratoire Ingénierie des Matériaux Polymères, CNRS UMR5223, Université Claude Bernard Lyon 1, F-69621 Villeurbanne, France. Electronic address: marie-therese.charreyre@ens-lyon.fr.
authors: Marie-Thérèse
authors: MT
authors: Charreyre
mesh_terms: D065638:Absorption, Radiation; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D016923:Cell Death; D002470:Cell Survival; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D004705:Endocytosis; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D019169:Jurkat Cells; D007700:Kinetics; D008856:Microscopy, Fluorescence; D038761:Photobleaching; D017785:Photons; D011108:Polymers / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D019265:Spectroscopy, Near-Infrared; D015027:Zebrafish / Q000196:embryology*
chemical_list: D001672:Biocompatible Materials; D005456:Fluorescent Dyes; D011108:Polymers
keywords: Fluorescence confocal microscopy; Multifunctional copolymer; Near-infrared; Polymer nanocarrier; Two-photon absorption; Zebrafish imaging