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  Cryofixation during live‐imaging enables millisecond time‐correlated light and electron microscopy.

Fuest, M., Nocera, G. M., Modena, M. M., Riedel, D., Mejia, Y. X., & Burg, T. P. (2018). Cryofixation during live‐imaging enables millisecond time‐correlated light and electron microscopy. Journal of Microscopy, 272(2), 87-95. doi:10.1111/jmi.12747.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0001-E70A-0 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-520F-1
Genre: Journal Article

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 Creators:
Fuest, M.1, Author              
Nocera, G. M.1, Author              
Modena, M. M.1, Author              
Riedel, D.2, Author              
Mejia, Y. X.1, Author              
Burg, T. P.1, Author              
Affiliations:
1Research Group of Biological Micro- and Nanotechnology, MPI for Biophysical Chemistry, Max Planck Society, ou_578602              
2Facility for Electron Microscopy, MPI for biophysical chemistry, Max Planck Society, ou_578615              

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 Abstract: Correlating live‐cell imaging with electron microscopy is among the most promising approaches to relate dynamic functions of cells or small organisms to their underlying ultrastructure. The time correlation between light and electron micrographs, however, is limited by the sample handling and fixation required for electron microscopy. Current cryofixation methods require a sample transfer step from the light microscope to a dedicated instrument for cryofixation. This transfer step introduces a time lapse of one second or more between live imaging and the fixed state, which is studied by electron microscopy. In this work, we cryofix Caenorhabditis elegans directly within the light microscope field of view, enabling millisecond time‐correlated live imaging and electron microscopy. With our approach, the time‐correlation is limited only by the sample cooling rate. C. elegans was used as a model system to establish compatibility of in situ cryofixation and subsequent transmission electron microscopy (TEM). TEM images of in situ cryofixed C. elegans show that the ultrastructure of the sample was well preserved with this method. We expect that the ability to correlate live imaging and electron microscopy at the millisecond scale will enable new paradigms to study biological processes across length scales based on real‐time selection and arrest of a desired state.

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Language(s): eng - English
 Dates: 2018-08-082018-11
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1111/jmi.12747
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Title: Journal of Microscopy
Source Genre: Journal
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Pages: - Volume / Issue: 272 (2) Sequence Number: - Start / End Page: 87 - 95 Identifier: -