Novel imaging approach reveals important details about rare eye disease choroideremia CommsBio
inhalation and then eyes were enucleated. For the pigmented mouse, immediately after enucleation, the eye was embedded in optical cutting temperature compound, rapidly frozen using acetone , and then cryosectioned through the center of the eye . A single cryosection was collected on a Super-frost Plus microscope slide , vacuum dried, and mounted in Immu-Mount immediately before imaging.
For the pigmented sample, microscopy was performed using an Axio Imager.Z2 slide scanning epifluorescence microscope equipped with a 20X/0.8 Plan-Apochromat non-immersion objective , a high resolution ORCA-Flash4.
For the albino sample, microscopy was performed using either a custom-modified confocal microscope or a custom assembled AO microscope. For imaging using the SP8, samples were transferred to glass slides and covered with a 0.17 mm thick coverslip with the photoreceptor layer facing the coverslip. Silicone grease was applied surrounding the sample to control the gap between the slide and the coverslip, in addition to anchoring the coverslip. A high NA objective was used.
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