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Bortoletto F.,National institute for astrophysics | Bonoli C.,National institute for astrophysics | Panizzolo P.,Fondazione per la Ricerca Biomedica Avanzata | Mammano F.,Fondazione per la Ricerca Biomedica Avanzata | And 2 more authors.
Journal of Microscopy | Year: 2011

Optical fibres with their unique ability to transport light even in a coherent way (fibre bundles) and the possibility to build small volume optical pieces (Graded Index Fibres, GRIN) have a dominant role in the assembly of probes and objectives for microscopy applications requiring noninvasive and flexible operation in small and crowded spaces (in vivo microscopy, endoscopy, inspection). Nowadays, even complex observing procedures like confocal, two-photon and optical coherence tomography can be approached with fibres, making possible in vivo applications and also in place decision and processing. We present here a series of analytical simulations and practical tests made on an experimental GRIN fibre objective light fed through an adaptive optics system aimed to verify the practical possibility to correct a focalized beam of light. We intend this as a first step to the implementation of non-invasive probes making use of forthcoming optical devices (scanners, deformable mirrors) based on MEMS technology. © 2010 The Authors Journal of Microscopy © 2010 Royal Microscopical Society.

Bortoletto F.,Istituto di Astrofisica Spaziale e Fisica Cosmica | Bonoli C.,Istituto di Astrofisica Spaziale e Fisica Cosmica | Panizzolo P.,Fondazione per la Ricerca Biomedica Avanzata | Ciubotaru C.D.,Fondazione per la Ricerca Biomedica Avanzata | And 3 more authors.
PLoS ONE | Year: 2011

Graded Index (GRIN) rod microlenses are increasingly employed in the assembly of optical probes for microendoscopy applications. Confocal, two-photon and optical coherence tomography (OCT) based on GRIN optical probes permit in-vivo imaging with penetration depths into tissue up to the centimeter range. However, insertion of the probe can be complicated by the need of several alignment and focusing mechanisms along the optical path. Furthermore, resolution values are generally not limited by diffraction, but rather by optical aberrations within the endoscope probe and feeding optics. Here we describe a multiphoton confocal fluorescence imaging system equipped with a compact objective that incorporates a GRIN probe and requires no adjustment mechanisms. We minimized the effects of aberrations with optical compensation provided by a low-order electrostatic membrane mirror (EMM) inserted in the optical path of the confocal architecture, resulting in greatly enhanced image quality. © 2011 Bortoletto et al.

Crispino G.,Fondazione per la Ricerca Biomedica Avanzata | Crispino G.,University of Padua | Di Pasquale G.,U.S. National Institutes of Health | Scimemi P.,University of Padua | And 13 more authors.
PLoS ONE | Year: 2011

The deafness locus DFNB1 contains GJB2, the gene encoding connexin26 and GJB6, encoding connexin30, which appear to be coordinately regulated in the inner ear. In this work, we investigated the expression and function of connexin26 and connexin30 from postnatal day 5 to adult age in double transgenic Cx26 Sox10Cre mice, which we obtained by crossing connexin26 floxed mice with a deleter Sox10-Cre line. Cx26 Sox10Cre mice presented with complete connexin26 ablation in the epithelial gap junction network of the cochlea, whereas connexin30 expression was developmentally delayed; immunolabeling patterns for both connexins were normal in the cochlear lateral wall. In vivo electrophysiological measurements in Cx26 Sox10Cre mice revealed profound hearing loss accompanied by reduction of endocochlear potential, and functional experiments performed in postnatal cochlear organotypic cultures showed impaired gap junction coupling. Transduction of these cultures with a bovine adeno associated virus vector restored connexin26 protein expression and rescued gap junction coupling. These results suggest that restoration of normal connexin levels by gene delivery via recombinant adeno associated virus could be a way to rescue hearing function in DFNB1 mouse models and, in future, lead to the development of therapeutic interventions in humans. © 2011 Crispino et al.

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