Individual filamentous phage imaged by electron holography
An in-line electron hologram of an individual f1.K phage was recorded with a purpose-built low energy electron point source (LEEPS) microscope. Cryo-microscopic methods were employed to prepare the specimen so that a single phage could be presented to the coherent low energy electrons: An aqueous ph...
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ftreroch:oai:doc.rero.ch:318026 2023-05-15T16:41:01+02:00 Individual filamentous phage imaged by electron holography Stevens, Gregory Krüger, Michael Latychevskaia, Tatiana Lindner, Peter Plückthun, Andreas Fink, Hans-Werner 2018-06-18T17:53:55Z http://doc.rero.ch/record/318026/files/249_2011_Article_743.pdf eng eng http://doc.rero.ch/record/318026/files/249_2011_Article_743.pdf 2018 ftreroch 2023-02-16T17:32:05Z An in-line electron hologram of an individual f1.K phage was recorded with a purpose-built low energy electron point source (LEEPS) microscope. Cryo-microscopic methods were employed to prepare the specimen so that a single phage could be presented to the coherent low energy electrons: An aqueous phage suspension was applied to a thin carbon membrane with micro-machined slits. The membrane was rapidly cooled to freeze the remaining water as an amorphous ice sheet, which was then sublimated at low temperatures and pressures to leave individual free-standing phages suspended across slits. An image of a phage particle, depicted as the amplitude of the object wave, was reconstructed numerically from a digitized record of the hologram, obtained using 88eV coherent electrons. The reconstructed image shows a single phage suspended across a slit in a supporting carbon membrane, magnified by a factor of 100,000. The width and shape in the reconstructed image compared well with a TEM image of the same filament. It is thus possible to record and reconstruct electron holograms of an individual phage. The challenge now is to improve the resolution of reconstructed images obtained by this method and to extend these structural studies to other biological molecules Other/Unknown Material Ice Sheet RERO DOC Digital Library |
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Open Polar |
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RERO DOC Digital Library |
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ftreroch |
language |
English |
description |
An in-line electron hologram of an individual f1.K phage was recorded with a purpose-built low energy electron point source (LEEPS) microscope. Cryo-microscopic methods were employed to prepare the specimen so that a single phage could be presented to the coherent low energy electrons: An aqueous phage suspension was applied to a thin carbon membrane with micro-machined slits. The membrane was rapidly cooled to freeze the remaining water as an amorphous ice sheet, which was then sublimated at low temperatures and pressures to leave individual free-standing phages suspended across slits. An image of a phage particle, depicted as the amplitude of the object wave, was reconstructed numerically from a digitized record of the hologram, obtained using 88eV coherent electrons. The reconstructed image shows a single phage suspended across a slit in a supporting carbon membrane, magnified by a factor of 100,000. The width and shape in the reconstructed image compared well with a TEM image of the same filament. It is thus possible to record and reconstruct electron holograms of an individual phage. The challenge now is to improve the resolution of reconstructed images obtained by this method and to extend these structural studies to other biological molecules |
author |
Stevens, Gregory Krüger, Michael Latychevskaia, Tatiana Lindner, Peter Plückthun, Andreas Fink, Hans-Werner |
spellingShingle |
Stevens, Gregory Krüger, Michael Latychevskaia, Tatiana Lindner, Peter Plückthun, Andreas Fink, Hans-Werner Individual filamentous phage imaged by electron holography |
author_facet |
Stevens, Gregory Krüger, Michael Latychevskaia, Tatiana Lindner, Peter Plückthun, Andreas Fink, Hans-Werner |
author_sort |
Stevens, Gregory |
title |
Individual filamentous phage imaged by electron holography |
title_short |
Individual filamentous phage imaged by electron holography |
title_full |
Individual filamentous phage imaged by electron holography |
title_fullStr |
Individual filamentous phage imaged by electron holography |
title_full_unstemmed |
Individual filamentous phage imaged by electron holography |
title_sort |
individual filamentous phage imaged by electron holography |
publishDate |
2018 |
url |
http://doc.rero.ch/record/318026/files/249_2011_Article_743.pdf |
genre |
Ice Sheet |
genre_facet |
Ice Sheet |
op_relation |
http://doc.rero.ch/record/318026/files/249_2011_Article_743.pdf |
_version_ |
1766031457867792384 |