Abstract
Electron microscopy offers necessary precision for the characterization of peptide materials at the nanoscale. Analysis is typically performed for acellular material specimens, whereas measurements in more complex, cellular environments prompt additional considerations for sample processing. Herein, we describe a protocol for the ultramicrotomy analysis of peptide-treated bacterial and mammalian cells. An emphasis is made on cell analysis following peptide treatment, as opposed to peptide analysis in cells, and focuses on sample processing, including fixation and staining procedures, resin embedding, sectioning, and imaging. The application of the protocol is demonstrated for intracellular measurements using antimicrobial peptide materials.
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Acknowledgments
This work was supported by UK’s Industry Strategy Challenge Fund. We thank the Electron microscopy imaging centre of the University of Sussex, funded by the School of Life Sciences, the Wellcome Trust (095605/Z/11/A, 208348/Z/17/Z) and the RM Phillips Trust for their support & assistance in this work.
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Rey, S., Faruqui, N., Ryadnov, M.G. (2021). Ultramicrotomy Analysis of Peptide-Treated Cells. In: Ryadnov, M. (eds) Polypeptide Materials. Methods in Molecular Biology, vol 2208. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0928-6_17
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DOI: https://doi.org/10.1007/978-1-0716-0928-6_17
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