The phototransductive membrane disks of a vertebrate photoreceptor outer segment (OS) are highly susceptible to perturbations during preservation for electron microscopy. To optimize their preservation for nanostructural studies, such as with electron tomography (ET), we developed a protocol, using a combination of chemical and physical fixation approaches, including transcardiac perfusion, high-pressure freezing, and freeze-substitution.
Photoreceptor Electron tomography High-pressure freezing Freeze-substitution Disk membrane morphogenesis
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This study was supported by NIH grants R01EY24667, R01EY13408, and P30EY00331. We thank Ivo Atanasov for technical assistance and acknowledge the use of instruments at the Electron Imaging Center for Nanomachines, supported by UCLA and NIH grant S10RR23057.
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