Work

Biological serial block face scanning electron microscopy at improved z-resolution based on Monte Carlo model

Year: 2018

Type: article

Abstract: . In SBEM, electrons with 1-3 keV energies are incident on a specimen block, from which backscattered electron (BSE) images are collected with x, y resolution of 5-10 nm in the block-face plane, and successive layers are removed by an in situ ultramicrotome. Spatial resolution along the z-direction, however, is limited to around 25 nm by the minimum cutting thickness. To improve the z-resolution, we have extracted depth information from BSE images acquired at dual primary beam energies, using Monte Carlo simulations of electron scattering. The relationship between depth of stain and ratio of dual-energy BSE intensities enables us to determine 3D structure with a ×2 improvement in z-resolution. We demonstrate the technique by sub-slice imaging of hepatocyte membranes in liver tissue.

Source: Scientific Reports

DOI: 10.1038/s41598-018-31231-w

Authors: Q. He, M. Hsueh, G. Zhang, D. C. Joy, R. D. Leapman

Institutions: National Institutes of Health, National Institute of Biomedical Imaging and Bioengineering, Oak Ridge National Laboratory, Center for Nanophase Materials Sciences, University of Tennessee at Knoxville

Language: English

Topic: Advanced Electron Microscopy Techniques and Applications

Subfield: Structural Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,690

Funders: National Institutes of Health, National Heart, Lung, and Blood Institute, National Institute of Biomedical Imaging and Bioengineering