Deutsch
 
Hilfe Datenschutzhinweis Impressum
  DetailsucheBrowse

Datensatz

 
 
DownloadE-Mail
  Electron cryotomography of vitrified cells with a Volta phase plate

Fukuda, Y., Laugks, U., Lucic, V., Baumeister, W., & Danev, R. (2015). Electron cryotomography of vitrified cells with a Volta phase plate. JOURNAL OF STRUCTURAL BIOLOGY, 190(2), 143-154. doi:10.1016/j.jsb.2015.03.004.

Item is

Externe Referenzen

einblenden:

Urheber

einblenden:
ausblenden:
 Urheber:
Fukuda, Yoshiyuki1, Autor           
Laugks, Ulrike1, Autor           
Lucic, Vladan1, Autor           
Baumeister, Wolfgang1, Autor           
Danev, Radostin1, Autor           
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

Inhalt

einblenden:
ausblenden:
Schlagwörter: CRYOELECTRON TOMOGRAPHY; IN-SITU; MICROSCOPY; NEURONS; MITOCHONDRIA; DIMERS; VIRUSVolta phase plate; Phase contrast; Electron cryotomography; Focused ion beam; Primary cultured neurons;
 Zusammenfassung: Electron cryotomography provides a means of studying the three dimensional structure of pleomorphic objects, such as organelles or cells, with a resolution of 1-3 nm. A limitation in the study of radiation sensitive biological samples is the low signal-to-noise ratio of the tomograms which may obscure fine details. To overcome this limitation, the recently developed Volta phase plate (VPP) was applied in electron cryotomographic studies of a wide range of cellular structures, from magnetotactic bacteria to primary cultured neurons. The results show that the VPP improves contrast significantly and consequently the signal-to-noise ratio of the tomograms, moreover it avoids disturbing fringing artifacts typical for Zernike phase plates. The contrast improvement provided by the VPP was also confirmed in projection images of relatively thick (similar to 400 nm) samples. In order to investigate the respective contributions of the VPP and the energy filter, images acquired with different combinations of the two were compared. Zero-loss energy filtering reduced the background noise in thicker areas of the sample and improved the contrast of features such as poly-beta-hydroxybutyrate granules in magnetotactic bacteria, whereas the VPP provided an overall contrast improvement for all sample areas. After 3D reconstruction, tomograms acquired with the combination of a VPP and an energy filter showed structural features in neuronal processes with outstanding clarity. We also show that the VPP can be combined with focused ion beam milling to examine structures embedded deeply inside cells. Thus, we expect that VPP will become a standard element of the electron cryotomography workflow. (C) 2015 Elsevier Inc. All rights reserved.

Details

einblenden:
ausblenden:
Sprache(n): eng - English
 Datum: 2015
 Publikationsstatus: Erschienen
 Seiten: 12
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: ISI: 000353753900005
DOI: 10.1016/j.jsb.2015.03.004
 Art des Abschluß: -

Veranstaltung

einblenden:

Entscheidung

einblenden:

Projektinformation

einblenden:

Quelle 1

einblenden:
ausblenden:
Titel: JOURNAL OF STRUCTURAL BIOLOGY
Genre der Quelle: Zeitschrift
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA : ACADEMIC PRESS INC ELSEVIER SCIENCE
Seiten: - Band / Heft: 190 (2) Artikelnummer: - Start- / Endseite: 143 - 154 Identifikator: ISSN: 1047-8477