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

© 2020 Elsevier Inc. All rights reserved..

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.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:158

Enthalten in:

Methods in cell biology - 158(2020) vom: 16., Seite 163-181

Sprache:

Englisch

Beteiligte Personen:

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

Links:

Volltext

Themen:

3DEM
Biopsy
Cancer
Electron microscopy
FIB-SEM
Journal Article
Large area mapping
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
SBF-SEM
Tissue processing
Volume rendering

Anmerkungen:

Date Completed 08.07.2021

Date Revised 08.07.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1016/bs.mcb.2020.01.005

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM310062462