Preservation of Photoreceptor Nanostructure for Electron Tomography Using Transcardiac Perfusion Followed by High-Pressure Freezing and Freeze-Substitution

  • Stefanie VollandEmail author
  • David S. WilliamsEmail author
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1074)


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 



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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Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Ophthalmology, Stein Eye Institute, UCLA School of MedicineLos AngelesUSA
  2. 2.Department of NeurobiologyUCLA School of MedicineLos AngelesUSA
  3. 3.Molecular Biology Institute, UCLA School of MedicineLos AngelesUSA
  4. 4.Brain Research Institute, UCLA School of MedicineLos AngelesUSA

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