Electroretinograms and fundal imaging was performed as described in Findlay et al., 2018 (link). PCM1-SNAP retinal labeling was carried out under inhaled anesthesia. 1.5 μl of 0.6 μM SNAP-Cell 647-SiR (New England Biolabs) was injected into the mouse vitreous under direct visualization using a Zeiss operating microscope. After 2 hr, mice were sacrificed by cervical dislocation and eyes enucleated. Keratectomy, sclerectomy and lensectomy were performed and whole retinas isolated. Flat mount petaloid retinal explants were made and mounted, photoreceptor side up, on Menzel_Glaser Superfrost Plus Gold slides (Thermo Fisher Scientific; K5800AMNZ72). Nuclei were stained with DAPI and mounted in Prolong Gold under coverslip. Slices were imaged on an Andor Dragonfly spinning disc confocal.
Other organizations :
MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, University of California, San Francisco, Sinai Health System, Lunenfeld-Tanenbaum Research Institute, Universitat de València, Universitat Jaume I, Edinburgh Cancer Research, University of Toronto, Chan Zuckerberg Initiative (United States)
Injection of 1.5 μl of 0.6 μM SNAP-Cell 647-SiR into the mouse vitreous
dependent variables
Electroretinograms
Fundal imaging
PCM1-SNAP retinal labeling
control variables
Inhaled anesthesia during PCM1-SNAP retinal labeling
Cervical dislocation and eye enucleation after 2 hr
Keratectomy, sclerectomy and lensectomy to isolate whole retinas
Flat mount petaloid retinal explants mounted photoreceptor side up on slides
Nuclei stained with DAPI and mounted in Prolong Gold under coverslip
Imaging on an Andor Dragonfly spinning disc confocal
Annotations
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