Live imaging of mammalian retina: Rod outer segments are stained by conventional mitochondrial dyes

Paolo Bianchini, Daniela Calzia, Silvia Ravera, Giovanni Candiano, Angela Bachi, Alessandro Morelli, Maurizio Bruschi, Isidoro M. Pepe, Alberto Diaspro, Isabella Panfoli

Research output: Contribution to journalArticle

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.

Original languageEnglish
Article number054017
JournalJournal of Biomedical Optics
Volume13
Issue number5
DOIs
Publication statusPublished - 2008

Keywords

  • confocal laser scanning microscopy
  • fluorescent dyes
  • imaging
  • JC-1
  • mitochondrial probes
  • MitoTracker
  • retina

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics

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    Bianchini, P., Calzia, D., Ravera, S., Candiano, G., Bachi, A., Morelli, A., Bruschi, M., Pepe, I. M., Diaspro, A., & Panfoli, I. (2008). Live imaging of mammalian retina: Rod outer segments are stained by conventional mitochondrial dyes. Journal of Biomedical Optics, 13(5), [054017]. https://doi.org/10.1117/1.2982528