RT Journal Article
SR Electronic
T1 A workflow for visualizing human cancer biopsies using large-format electron microscopy
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 675371
DO 10.1101/675371
A1 Jessica L. Riesterer
A1 Claudia S. López
A1 Erin S. Stempinski
A1 Melissa Williams
A1 Kevin Loftis
A1 Kevin Stoltz
A1 Guillaume Thibault
A1 Christian Lanicault
A1 Todd Williams
A1 Joe W. Gray
YR 2019
UL http://biorxiv.org/content/early/2019/06/19/675371.abstract
AB 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.