TY  - JOUR
A1  - Perkovic, Mario
A1  - Kunz, Michael
A1  - Endesfelder, Ulrike
A1  - Bunse, Stefanie
A1  - Wigge, Christoph
A1  - Yu, Zhou
A1  - Hodirnau, Victor-Valentin
A1  - Scheffer, Margot
A1  - Seybert, Anja
A1  - Malkusch, Sebastian
A1  - Schuman, Erin
A1  - Heilemann, Mike
A1  - Frangakis, Achilleas S.
T1  - Correlative light- and electron microscopy with chemical tags
T2  - Journal of structural biology
N2  - 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.
KW  - Correlative electron and light microscopy
KW  - Electron tomography
KW  - Adherens junctions
Y1  - 2014
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/34811
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-348115
SN  - 1095-8657
SN  - 1047-8477
N1  - This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).
VL  - 186
IS  - 2
SP  - 205
EP  - 213
PB  - Elsevier
CY  - San Diego, Calif.
ER  -