Abstract
Contemporarily, serial block-face scanning electron microscopy (SBF-SEM) has emerged as an immensely powerful nanoscopic imaging technique, capable of generating large-volume three-dimensional information on a variety of biological specimens in a semiautomated manner. Despite the plethora of insights and advantages provided by SBF-SEM, a major challenge inherent to the technique is that of electron charging, which ultimately reduces attainable resolution and detracts from overall image quality. In this chapter, we describe a pre-embedding approach that involves infiltration of tissue with a highly conductive silver filler suspension following primary fixation. Such an approach is demonstrated to improve overall sample conductivity, resulting in the minimization of charging under high-vacuum conditions and an improvement in lateral resolution and image contrast. The strength of this sample preparation approach for SBF-SEM is illustrated on liver tissue.
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Acknowledgments
The authors acknowledge the facilities as well as the technical assistance from staff of the AMMRF node at the University of Sydney. We are also grateful to Dr. Laura A. Lindsay and Professor Christopher R. Murphy from the Cell and Reproductive Biology Laboratory, the University of Sydney, for their kind provision of tissue samples.
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Shami, G.J., Cheng, D., Braet, F. (2018). Silver Filler Pre-embedding to Enhance Resolution and Contrast in Multidimensional SEM: A Nanoscale Imaging Study on Liver Tissue. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_33
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DOI: https://doi.org/10.1007/978-1-4939-8591-3_33
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