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Verfasst von:Schorb, Martin [VerfasserIn]   i
 Bykov, Yury [VerfasserIn]   i
 Briggs, John A. G. [VerfasserIn]   i
Titel:New hardware and workflows for semi-automated correlative cryo-fluorescence and cryo-electron microscopy/tomography
Verf.angabe:Martin Schorb, Leander Gaechter, Ori Avinoam, Frank Sieckmann, Mairi Clarke, Cecilia Bebeacua, Yury S. Bykov, Andreas F.-P. Sonnen, Reinhard Lihl, John A.G. Briggs
Jahr:2017
Jahr des Originals:2016
Umfang:11 S.
Teil:volume:197
 year:2017
 number:2
 pages:83-93
 extent:11
Fussnoten:Published online: 28 June 2016 ; Gesehen am 27.07.2018
Titel Quelle:Enthalten in: Journal of structural biology
Ort Quelle:San Diego, Calif. : Elsevier, 1990
Jahr Quelle:2017
Band/Heft Quelle:197(2017), 2, Seite 83-93
ISSN Quelle:1095-8657
Abstract:Correlative light and electron microscopy allows features of interest defined by fluorescence signals to be located in an electron micrograph of the same sample. Rare dynamic events or specific objects can be identified, targeted and imaged by electron microscopy or tomography. To combine it with structural studies using cryo-electron microscopy or tomography, fluorescence microscopy must be performed while maintaining the specimen vitrified at liquid-nitrogen temperatures and in a dry environment during imaging and transfer. Here we present instrumentation, software and an experimental workflow that improves the ease of use, throughput and performance of correlated cryo-fluorescence and cryo-electron microscopy. The new cryo-stage incorporates a specially modified high-numerical aperture objective lens and provides a stable and clean imaging environment. It is combined with a transfer shuttle for contamination-free loading of the specimen. Optimized microscope control software allows automated acquisition of the entire specimen area by cryo-fluorescence microscopy. The software also facilitates direct transfer of the fluorescence image and associated coordinates to the cryo-electron microscope for subsequent fluorescence-guided automated imaging. Here we describe these technological developments and present a detailed workflow, which we applied for automated cryo-electron microscopy and tomography of various specimens.
DOI:doi:10.1016/j.jsb.2016.06.020
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext ; Verlag: http://dx.doi.org/10.1016/j.jsb.2016.06.020
 Volltext: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5287355/
 DOI: https://doi.org/10.1016/j.jsb.2016.06.020
Datenträger:Online-Ressource
Sprache:eng
K10plus-PPN:1578007291
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