Format

Send to

Choose Destination
J Struct Biol. 2019 Jul 1;207(1):1-11. doi: 10.1016/j.jsb.2019.03.008. Epub 2019 Mar 23.

Semi-automated 3D segmentation of human skeletal muscle using Focused Ion Beam-Scanning Electron Microscopic images.

Author information

1
University of British Columbia, Vancouver, Canada.
2
Longitudinal Studies Section, National Institute on Aging, National Institutes of Health, Baltimore, MD 21225, USA.
3
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
4
Longitudinal Studies Section, National Institute on Aging, National Institutes of Health, Baltimore, MD 21225, USA. Electronic address: luigi.ferrucci@nih.gov.
5
University of British Columbia, Vancouver, Canada. Electronic address: Sriram.Subramaniam@ubc.ca.

Abstract

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) is an imaging approach that enables analysis of the 3D architecture of cells and tissues at resolutions that are 1-2 orders of magnitude higher than that possible with light microscopy. The slow speeds of data collection and manual segmentation are two critical problems that limit the more extensive use of FIB-SEM technology. Here, we present an easily accessible robust method that enables rapid, large-scale acquisition of data from tissue specimens, combined with an approach for semi-automated data segmentation using the open-source machine learning Weka segmentation software, which dramatically increases the speed of image analysis. We demonstrate the feasibility of these methods through the 3D analysis of human muscle tissue by showing that our process results in an improvement in speed of up to three orders of magnitude as compared to manual approaches for data segmentation. All programs and scripts we use are open source and are immediately available for use by others.

KEYWORDS:

3D electron microscopy; Aging; FIB-SEM; Machine learning; Mitochondrial structure; Semi-automated segmentation; Skeletal muscle; Tissue imaging

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center