title: Organization of inner cellular components as reported by a viscosity-sensitive fluorescent Bodipy probe suitable for phasor approach to FLIM.
abstract: According to the recent developments in imaging strategies and in tailoring fluorescent molecule as probe for monitoring biological systems, we coupled a Bodipy-based molecular rotor (BoMe) with FLIM phasor approach to evaluate the viscosity in different intracellular domains. BoMe rapidly permeates cells, stains cytoplasmic as well as nuclear domains, and its optical properties make it perfectly suited for widely diffused confocal microscopy imaging setups. The capability of BoMe to report on intracellular viscosity was put to the test by using a cellular model of a morbid genetic pathology (Hutchinson-Gilford progeria syndrome, HGPS). Our results show that the nucleoplasm of HGPS cells display reduced viscosity as compared to normal cells. Since BoMe displays significant affinity towards DNA, as demonstrated by an in vitro essay, we hypothesize that genetic features of HGPS, namely the misassembly of lamin A protein within the nuclear lamina, modulates chromatin compaction. This hypothesis nicely agrees with literature data.
authors.affiliation: NEST, Scuola Normale Superiore and NANO-CNR, Pisa, Italy.
authors.forename: Gianmarco
authors.initials: G
authors.lastname: Ferri
authors.affiliation: Center for Nanotechnology Innovation @ NEST, Istituto Italiano di Tecnologia, Pisa, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Nucara
authors.affiliation: Dipartimento di Chimica e Chimica Industriale, Università di Pisa, 56124 Pisa, Italy.
authors.forename: Tarita
authors.initials: T
authors.lastname: Biver
authors.affiliation: NEST, Scuola Normale Superiore and NANO-CNR, Pisa, Italy; Istituto di Biofisica - CNR, via Ugo la Malfa 153, 90146 Palermo, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Battisti
authors.affiliation: Center for Nanotechnology Innovation @ NEST, Istituto Italiano di Tecnologia, Pisa, Italy. Electronic address: giovanni.signore@iit.it.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Signore
authors.affiliation: NEST, Scuola Normale Superiore and NANO-CNR, Pisa, Italy; Istituto di Biofisica - CNR, via Moruzzi 1, 56124 Pisa, Italy. Electronic address: ranieri.bizzarri@nano.cnr.it.
authors.forename: Ranieri
authors.initials: R
authors.lastname: Bizzarri
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000737:chemistry*; D002417:Cattle; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D015394:Molecular Structure; D011371:Progeria / Q000235:genetics / Q000473:pathology*; D014783:Viscosity
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D004247:DNA; C427203:calf thymus DNA
keywords: Chromatin compaction; FLIM; Hutchinson–Guilford progeria syndrome; Intracellular viscosity; Molecular rotor; Phasor approach