A workflow for visualizing human cancer biopsies using large-format electron microscopy

Jessica L. Riesterer, Claudia S. López, Erin S. Stempinski, Melissa Williams, Kevin Loftis, Kevin Stoltz, Guillaume Thibault, Christian Lanicault, Todd Williams, Joe W. Gray

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Recent developments in large format electron microscopy have enabled generation of images that provide detailed ultrastructural information on normal and diseased cells and tissues. Analyses of these images increase our understanding of cellular organization and interactions and disease-related changes therein. In this manuscript, we describe a workflow for two-dimensional (2D) and three-dimensional (3D) imaging, including both optical and scanning electron microscopy (SEM) methods, that allow pathologists and cancer biology researchers to identify areas of interest from human cancer biopsies. The protocols and mounting strategies described in this workflow are compatible with 2D large format EM mapping, 3D focused ion beam-SEM and serial block face-SEM. The flexibility to use diverse imaging technologies available at most academic institutions makes this workflow useful and applicable for most life science samples. Volumetric analysis of the biopsies studied here revealed morphological, organizational and ultrastructural aspects of the tumor cells and surrounding environment that cannot be revealed by conventional 2D EM imaging. Our results indicate that although 2D EM is still an important tool in many areas of diagnostic pathology, 3D images of ultrastructural relationships between both normal and cancerous cells, in combination with their extracellular matrix, enables cancer researchers and pathologists to better understand the progression of the disease and identify potential therapeutic targets.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
EditorsPhong Tran
PublisherAcademic Press Inc.
Pages163-181
Number of pages19
ISBN (Print)9780128200087
DOIs
StatePublished - 2020

Publication series

NameMethods in Cell Biology
Volume158
ISSN (Print)0091-679X

Keywords

  • 3DEM
  • Biopsy
  • Cancer
  • Electron microscopy
  • FIB-SEM
  • Large area mapping
  • SBF-SEM
  • Tissue processing
  • Volume rendering

ASJC Scopus subject areas

  • Cell Biology

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  • Cite this

    Riesterer, J. L., López, C. S., Stempinski, E. S., Williams, M., Loftis, K., Stoltz, K., Thibault, G., Lanicault, C., Williams, T., & Gray, J. W. (2020). A workflow for visualizing human cancer biopsies using large-format electron microscopy. In P. Tran (Ed.), Methods in Cell Biology (pp. 163-181). (Methods in Cell Biology; Vol. 158). Academic Press Inc.. https://doi.org/10.1016/bs.mcb.2020.01.005