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Low-Temperature Electron Microscopy: Techniques and Protocols

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Cryopreservation and Freeze-Drying Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1257))

Abstract

Low-temperature electron microscopy endeavors to provide “solidification of a biological specimen by cooling with the aim of minimal displacement of its components through the use of low temperature as a physical fixation strategy” (Steinbrecht and Zierold, Cryotechniques in biological electron microscopy. Springer-Verlag, Berlin, p 293, 1987). The intention is to maintain confidence that the tissue observed retains the morphology and dimensions of the living material while also ensuring soluble cellular components are not displaced. As applied to both scanning and transmission electron microscopy, cryo-electron microscopy is a strategy whereby the application of low-temperature techniques are used to reduce or remove processing artifacts which are commonly encountered in more conventional room temperature electron microscopy techniques which rely heavily on chemical fixation and heavy metal staining. Often, cryo-electron microscopy allows direct observation of specimens, which have not been stained or chemically fixed.

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Acknowledgments

Kirsty MacLellan-Gibson who has helped organize and run the NIBSC cryoworkshop, the team at CUI, Leanne Glover, Gema Vizcay, Monika Balys, Fiona Winning, and Pippa Hawes (The Pirbright Institute) for her helpful suggestions and comments.

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Authors and Affiliations

  1. Centre for Ultrastructural Imaging, King’s College London, New Hunts House, Guy’s Campus, London, SE1 1UL, UK

    Roland A. Fleck Ph.D.

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  1. Roland A. Fleck Ph.D.
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Correspondence to Roland A. Fleck Ph.D. .

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Editors and Affiliations

  1. Institute of Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany

    Willem F. Wolkers

  2. Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany

    Harriëtte Oldenhof

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Fleck, R.A. (2015). Low-Temperature Electron Microscopy: Techniques and Protocols. In: Wolkers, W., Oldenhof, H. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology, vol 1257. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-2193-5_9

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  • DOI: https://doi.org/10.1007/978-1-4939-2193-5_9

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-2192-8

  • Online ISBN: 978-1-4939-2193-5

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