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

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Retinal Degenerative Diseases

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1074))

Abstract

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.

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References

  • McDonald KL, Auer M (2006) High-pressure freezing, cellular tomography, and structural cell biology. BioTechniques 4:137–143. 139, 141

    Article  Google Scholar 

  • Murk JL, Posthuma G, Koster AJ et al (2003) Influence of aldehyde fixationon on the morphology of endosomes and lysosomes: quantitative analysis and electron tomohgraphy. J Microsc 212:81–90

    Article  CAS  Google Scholar 

  • Sosinsky GE, Crum J, Jones YJ et al (2008) The combination of chemical fixation procedures with high pressure freezing and freeze substitution preserves highly labile tissue ultrastructure for electron tomography applications. J Struct Biol 161:359–371

    Article  CAS  Google Scholar 

  • Studer D, Graber W, Al-Amoudi A, Eggli P (2001) A new approach for cryofixation by high-pressure freezing. J Microsc 203:285–294

    Article  CAS  Google Scholar 

  • Volland S, Louise CH, Kong C et al (2015) Three-dimensional organization of nascent rod outer segment disk membranes. PNAS 112:14870–14875

    Article  CAS  Google Scholar 

  • Young RW (1967) The renewal of photoreceptor cell outer segments. J Cell Biol 33:61–72

    Article  CAS  Google Scholar 

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Acknowledgments

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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Authors and Affiliations

  1. Department of Ophthalmology, Stein Eye Institute, UCLA School of Medicine, Los Angeles, CA, USA

    Stefanie Volland & David S. Williams

  2. Department of Neurobiology, UCLA School of Medicine, Los Angeles, CA, USA

    David S. Williams

  3. Molecular Biology Institute, UCLA School of Medicine, Los Angeles, CA, USA

    David S. Williams

  4. Brain Research Institute, UCLA School of Medicine, Los Angeles, CA, USA

    David S. Williams

Authors
  1. Stefanie Volland
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  2. David S. Williams
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Corresponding authors

Correspondence to Stefanie Volland or David S. Williams .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, University of Florida, Gainesville, Florida, USA

    John D. Ash

  2. Ophthalmology and Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

    Robert E. Anderson

  3. School of Medicine Beckman Vision Center, University of California San Francisco, San Francisco, California, USA

    Matthew M. LaVail

  4. Departments of Ophthalmology and Cell Biology, Duke Eye Center, Duke University Medical Center, Durham, North Carolina, USA

    Catherine Bowes Rickman

  5. Division of Ophthalmology, Case Western Reserve University, Cleveland, Ohio, USA

    Joe G. Hollyfield

  6. Department of Ophthalmology, University Hospital Zurich, Zurich, Switzerland

    Christian Grimm

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Volland, S., Williams, D.S. (2018). Preservation of Photoreceptor Nanostructure for Electron Tomography Using Transcardiac Perfusion Followed by High-Pressure Freezing and Freeze-Substitution. In: Ash, J., Anderson, R., LaVail, M., Bowes Rickman, C., Hollyfield, J., Grimm, C. (eds) Retinal Degenerative Diseases. Advances in Experimental Medicine and Biology, vol 1074. Springer, Cham. https://doi.org/10.1007/978-3-319-75402-4_73

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