Document Detail

Live imaging of mammalian retina: rod outer segments are stained by conventional mitochondrial dyes.
MedLine Citation:
PMID:  19021397     Owner:  NLM     Status:  MEDLINE    
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.
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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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of biomedical optics     Volume:  13     ISSN:  1083-3668     ISO Abbreviation:  J Biomed Opt     Publication Date:    2008 Sep-Oct
Date Detail:
Created Date:  2008-11-21     Completed Date:  2009-02-27     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9605853     Medline TA:  J Biomed Opt     Country:  United States    
Other Details:
Languages:  eng     Pagination:  054017     Citation Subset:  IM    
University of Genoa, Laboratory for Advanced, Microscopy, Bioimaging, and Spectroscopy (LAMBS), MicroSCoBiO Research Center, Department of Physics, Via Dodecaneso 33, 16146, Genoa, Italy.
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MeSH Terms
Contrast Media
Fluorescent Dyes*
Image Enhancement / methods*
Microscopy, Confocal / methods*
Retinal Rod Photoreceptor Cells / cytology*
Retinoscopy / methods*
Reg. No./Substance:
0/Aldehydes; 0/Benzimidazoles; 0/Carbocyanines; 0/Contrast Media; 0/Fluorescent Dyes; 0/mitotracker green FM; 21527-78-6/5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine

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