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  Correlative STED super-resolution light and electron microscopy on resin sections.

Wurm, C. A., Schwarz, H., Jans, D. C., Riedel, D., Humbel, B. M., & Jakobs, S. (2019). Correlative STED super-resolution light and electron microscopy on resin sections. Journal of Physics D: Applied Physics, 52(37): 374003. doi:10.1088/1361-6463/ab2b31.

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 Creators:
Wurm, C. A.1, Author           
Schwarz, H., Author
Jans, D. C.2, Author                 
Riedel, D.3, Author           
Humbel, B. M., Author
Jakobs, S.2, Author           
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              
2Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              
3Facility for Electron Microscopy, MPI for biophysical chemistry, Max Planck Society, ou_578615              

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Free keywords: CLEM; STED; super-resolution microscopy; immunolabelling; immunogold; resin embedding; correlative microscopy
 Abstract: Correlative light and electron microscopy approaches can reveal the localisation of specific proteins while providing detailed information on the cellular context, thereby combining the strengths of both imaging modalities. The major challenge in combining fluorescence microscopy with electron microscopy is the different sample preparation requirements necessary for obtaining high quality data from both modalities. To overcome this limitation, we combined conventional sample preparation protocols for electron microscopy with post-embedding labelling on ultra-thin sections using antibodies and other specific ligands. We successfully employed STED super-resolution microscopy to image the subcellular distribution of several targets in various specimen including E. coli, T brucei, S. cerevisiae, human cancer cells and bovine sperm. Thus, we present widely applicable methods facilitating the use of antibodies for correlative super-resolution light and electron microscopy of post-embedding labelled targets.

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Language(s): eng - English
 Dates: 2019-07-162019-09-11
 Publication Status: Issued
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 Rev. Type: Peer
 Identifiers: DOI: 10.1088/1361-6463/ab2b31
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Title: Journal of Physics D: Applied Physics
Source Genre: Journal
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Pages: 8 Volume / Issue: 52 (37) Sequence Number: 374003 Start / End Page: - Identifier: -