Journal
JOURNAL OF BIOMEDICAL OPTICS
Volume 13, Issue 5, Pages -Publisher
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.2982528
Keywords
confocal laser scanning microscopy; fluorescent dyes; mitochondrial probes; retina; imaging; JC-1; MitoTracker
Funding
- Italian Ministry for Research and Technology (MIUR)
- Istituto Firc di Oncologia Morelcolare (IFOM), Milan
- Italian Research Program of National Interest [2006028909]
- Fondazione San Paolo (Torino, Italy)
- RenalChild Foundation
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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. c 2008 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.2982528]
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