Mitotracker Deep Red is a far-red fluorescent dye that chemically stains mitochondria in live cells (Audano et al., 2021 (link); Weiss-Sadan et al., 2019 (link)). Fresh 200 μM of MitoTracker® Deep Red FM (Cell Signaling Technology, #8778) dye solution was prepared by diluting a 5 mM of stock solution prepared in DMSO with filtered 0.5X MMR on the day of injection. The Mitotracker solution was front loaded into a borosilicate glass capillary (World Precision Instruments) that had been pulled and then broken to 1–2 μm tip diameter using fine forceps, and then injected intravitreally into left eyes of anesthetized animals at around NF stage 48 (Nieuwkoop and Faber, 1994 ), using a pressure injector (Narishige IM-300 Microinjector); successful injections were verified by observing an acute but mild swelling of the eye at the time of injection. After injection, the animals were returned to a new dish containing 0.1X MMR at room temperature and imaged 3.5 hours after injection for S1A and S1B and 15–18 hours after injection for all the other experiments.
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Intravitreal Injection of Mitotracker Deep Red
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Corresponding Organization : University of California, Davis
Other organizations : Stanford University, University of California, San Diego
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Variable analysis
independent variables
- Injection of Mitotracker Deep Red dye solution
- Mitochondrial staining in live cells
- Anesthetized animals at around NF stage 48
- Injection into left eyes of animals
- Imaging at 3.5 hours and 15-18 hours after injection
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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