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  Aberration-corrected cryoimmersion light microscopy.

Faoro, R., Bassu, M., Mejia, Y. X., Stephan, T., Dudani, N., Boeker, C., et al. (2018). Aberration-corrected cryoimmersion light microscopy. Proceedings of the National Academy of Sciences of the United States of America, 115(6), 1204-1209. doi:10.1073/pnas.1717282115.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0000-33ED-C Version Permalink: http://hdl.handle.net/21.11116/0000-0003-4F56-5
Genre: Journal Article

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 Creators:
Faoro, R.1, Author              
Bassu, M.1, Author              
Mejia, Y. X.1, Author              
Stephan, T.2, Author              
Dudani, N., Author
Boeker, C., Author
Jakobs, S.2, Author              
Burg, T. P.1, Author              
Affiliations:
1Research Group of Biological Micro- and Nanotechnology, MPI for Biophysical Chemistry, Max Planck Society, ou_578602              
2Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              

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 Abstract: Cryogenic fluorescent light microscopy of flash-frozen cells stands out by artifact-free fixation and very little photobleaching of the fluorophores used. To attain the highest level of resolution, aberration-free immersion objectives with accurately matched immersion media are required, but both do not exist for imaging below the glass-transition temperature of water. Here, we resolve this challenge by combining a cryoimmersion medium, HFE-7200, which matches the refractive index of room-temperature water, with a technological concept in which the body of the objective and the front lens are not in thermal equilibrium. We implemented this concept by replacing the metallic front-lens mount of a standard bioimaging water immersion objective with an insulating ceramic mount heated around its perimeter. In this way, the objective metal housing can be maintained at room temperature, while creating a thermally shielded cold microenvironment around the sample and front lens. To demonstrate the range of potential applications, we show that our method can provide superior contrast in Escherichia coli and yeast cells expressing fluorescent proteins and resolve submicrometer structures in multicolor immunolabeled human bone osteosarcoma epithelial (U2OS) cells at −140°C.

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Language(s): eng - English
 Dates: 2018-01-22
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1073/pnas.1717282115
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Title: Proceedings of the National Academy of Sciences of the United States of America
Source Genre: Journal
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Pages: - Volume / Issue: 115 (6) Sequence Number: - Start / End Page: 1204 - 1209 Identifier: -