Correlative light- and electron microscopy with chemical tags

  • Correlative microscopy incorporates the specificity of fluorescent protein labeling into high-resolution electron micrographs. Several approaches exist for correlative microscopy, most of which have used the green fluorescent protein (GFP) as the label for light microscopy. Here we use chemical tagging and synthetic fluorophores instead, in order to achieve protein-specific labeling, and to perform multicolor imaging. We show that synthetic fluorophores preserve their post-embedding fluorescence in the presence of uranyl acetate. Post-embedding fluorescence is of such quality that the specimen can be prepared with identical protocols for scanning electron microscopy (SEM) and transmission electron microscopy (TEM); this is particularly valuable when singular or otherwise difficult samples are examined. We show that synthetic fluorophores give bright, well-resolved signals in super-resolution light microscopy, enabling us to superimpose light microscopic images with a precision of up to 25 nm in the x-y plane on electron micrographs. To exemplify the preservation quality of our new method we visualize the molecular arrangement of cadherins in adherens junctions of mouse epithelial cells.

Download full text files

Export metadata

Additional Services

Share in Twitter Search Google Scholar
Author:Mario Perkovic, Michael Kunz, Ulrike Endesfelder, Stefanie Bunse, Christoph Wigge, Zhou Yu, Victor-Valentin Hodirnau, Margot P. Scheffer, Anja Seybert, Sebastian Malkusch, Erin Schuman, Mike HeilemannORCiDGND, Achilleas S. Frangakis
Pubmed Id:
Parent Title (English):Journal of structural biology
Place of publication:San Diego, Calif.
Document Type:Article
Year of Completion:2014
Date of first Publication:2014/03/31
Publishing Institution:Universit├Ątsbibliothek Johann Christian Senckenberg
Release Date:2014/10/27
Tag:Adherens junctions; Correlative electron and light microscopy; Electron tomography
Page Number:9
First Page:205
Last Page:213
This is an open access article under the CC BY-NC-ND license (
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Angeschlossene und kooperierende Institutionen / MPI f├╝r Hirnforschung
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
Licence (German):License LogoCreative Commons - Namensnennung-Nicht kommerziell-Keine Bearbeitung 3.0