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Journal of Biomedical Optics

Live imaging of mammalian retina: rod outer segments are stained by conventional mitochondrial dyes
Author(s): Paolo Bianchini; Daniela Calzia; Silvia Ravera; Giovanni Candiano; Angela Bachi; Alessandro Morelli; Maurizio Bruschi; Isidoro M. Pepe; Alberto Diaspro; Isabella Panfoli
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Paper Abstract

The vertebrate retina is an array of “narrow-capture” photoreceptive elements of diverse cellular types that allow the fine spatial resolution characteristic of vision. Imaging of photoreceptors and of the whole retina has been previously reported; however, both were achieved exclusively after fixation. We report our development of a new technique for imaging live bovine retinas ex vivo. Using this technique, we conducted fluorescence confocal laser scanning microscopic imaging of bovine retinas. Eyecups were incubated with conventional fluorescent mitochondrial probes (MitoTracker and JC-1). Unexpectedly, we found that, besides the retinal mitochondria, the rod outer segments that are devoid of mitochondria were also stained. No other neuron was stained. Both protonophores, which decrease mitochondrial membrane potential, or inhibit electron transport strongly inhibited the selective association of dyes with both retinal rod outer segments and mitochondria. This is the first time that living rod outer segments were visualized by this technique. This finding may shed light on previous reports of the existence of a proton potential across the disk membranes and on the mechanism of the adenosine tri-phosphate (ATP) supply for phototransduction, which still requires investigation.

Paper Details

Date Published: 1 September 2008
PDF: 6 pages
J. Biomed. Opt. 13(5) 054017 doi: 10.1117/1.2982528
Published in: Journal of Biomedical Optics Volume 13, Issue 5
Show Author Affiliations
Paolo Bianchini, Univ. degli Studi di Genova (Italy)
Daniela Calzia, Univ. degli Studi di Genova (Italy)
Silvia Ravera, Univ. degli Studi di Genova (Italy)
Giovanni Candiano, G. Gaslini Children's Hospital (Italy)
Angela Bachi, Fondazione San Raffaele del Monte Tabor (Italy)
Alessandro Morelli, Univ. degli Studi di Genova (Italy)
Maurizio Bruschi, G. Gaslini Children's Hospital (Italy)
Isidoro M. Pepe, Univ. degli Studi di Genova (Italy)
Alberto Diaspro, Univ. degli Studi di Genova (Italy)
Isabella Panfoli, Univ. degli Studi di Genova (Italy)

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