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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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Faoro, R.1, Autor           
Bassu, M.1, Autor           
Mejia, Y. X.1, Autor           
Stephan, T.2, Autor           
Dudani, N., Autor
Boeker, C., Autor
Jakobs, S.2, Autor           
Burg, T. P.1, Autor           
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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 Zusammenfassung: 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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Sprache(n): eng - English
 Datum: 2018-01-22
 Publikationsstatus: Online veröffentlicht
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 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1073/pnas.1717282115
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Titel: Proceedings of the National Academy of Sciences of the United States of America
Genre der Quelle: Zeitschrift
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Seiten: - Band / Heft: 115 (6) Artikelnummer: - Start- / Endseite: 1204 - 1209 Identifikator: -