Digitala Vetenskapliga Arkivet

Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Laboratory cryo x-ray microscopy for 3D cell imaging
KTH, School of Engineering Sciences (SCI), Applied Physics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Biomedical and X-ray Physics.ORCID iD: 0000-0002-3717-7307
KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.ORCID iD: 0000-0001-5178-7593
Show others and affiliations
2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, article id 13433Article in journal (Refereed) Published
Abstract [en]

Water-window x-ray microscopy allows two-and three-dimensional (2D and 3D) imaging of intact unstained cells in their cryofixed near-native state with unique contrast and high resolution. Present operational biological water-window microscopes are based at synchrotron facilities, which limits their accessibility and integration with complementary methods. Laboratory-source microscopes have had difficulty addressing relevant biological tasks with proper resolution and contrast due to long exposure times and limited up-time. Here we report on laboratory cryo x-ray microscopy with the exposure time, contrast, and reliability to allow for routine high-spatial resolution 3D imaging of intact cells and cell-cell interactions. Stabilization of the laser-plasma source combined with new optics and sample preparation provide high-resolution cell imaging, both in 2D with ten-second exposures and in 3D with twenty-minute tomography. Examples include monitoring of the distribution of carbon-dense vesicles in starving HEK293T cells and imaging the interaction between natural killer cells and target cells.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP , 2017. Vol. 7, article id 13433
National Category
Physical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-217434DOI: 10.1038/s41598-017-13538-2ISI: 000413188400014PubMedID: 29044158Scopus ID: 2-s2.0-85031924153OAI: oai:DiVA.org:kth-217434DiVA, id: diva2:1158157
Note

QC 20171117

Available from: 2017-11-17 Created: 2017-11-17 Last updated: 2020-05-25Bibliographically approved
In thesis
1. Biological Laboratory X-Ray Microscopy
Open this publication in new window or tab >>Biological Laboratory X-Ray Microscopy
2020 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Soft x-ray microscopy in the water window (𝜆 ≈ 2.3 − 4.3 nm) is a powerful technique for high-resolution biological imaging. The strong natural contrast between carbon-based structures and water allows visualization of hydrated and unstained samples, while providing enough transmission through up to ∼ 10 μm of organic matter. Furthermore, the full potential of this technique can be exploited by performing computed tomography, thus obtaining a complete 3D image of the object.

Routine short-exposure water-window microscopy of whole cells and tissue is currently performed at synchrotron-radiation facilities around the world, but with a limited accessibility to the wider research community. For this reason, laboratory-based systems have been developed, which are now reaching maturity. The benefits compared to the synchrotron-based instruments include easier integration with complementary methods in the home laboratory, in addition to the increased access that allows for the often time-consuming optimization of experimental parameters as well as longitudinal studies.

This Thesis presents recent developments of the Stockholm laboratory x-ray microscope as well as several biological applications. Work has been done on improving the mechanical and thermal stability of the microscope, resulting in a resolution of 25 nm (half period) in images of test targets. The biological applications were enabled by a significantly increased x-ray flux through the system as well as an improved operational stability. This work demonstrates 10-second exposure imaging of whole cryofixed cells, imaging of viral infections in cells, and 20 minutes total exposure cryotomography.

Abstract [sv]

Röntgenmikroskopi i vattenfönstret (𝜆 ≈ 2.3−4.3 nm) är en kraftfull metodför högupplöst biologisk avbildning. Den naturligt höga kontrasten mellankolbaserade strukturer och vatten möjliggör visualisering av prover i ettnästintill opåverkat tillstånd, och ger samtidigt tillräcklig transmission genomupp till ∼ 10 μm organisk materia. Teknikens fulla potential utnyttjasvidare genom datortomografi, vilket resulterar i en fullständig 3D-bild avobjektet.Röntgenmikroskopi i vattenfönstret, av celler och vävnad och med kortexponeringstid, utförs rutinmässigt vid synkrotronljuskällor runt om i världen,men med begränsad tillgänglighet för forskarsamfundet. Av den anledningenhar laboratoriebaserade system utvecklats, vilka nu börjar nåmognad. Fördelarna jämfört med synkrotronbaserade instrument består avenklare integrering av komplementära laboratoriemetoder, utöver den utökadetillgängligheten som tillåter tidskrävande optimering av experimentellaparametrar såväl som longitudinella studier.Denna avhandling beskriver nyligen utfört arbete för att förbättra detkompakta mjukröntgenmikroskopet i Stockholm, samt flera biologiskatillämpningar. Arbete har gjorts för att förbättra mikroskopets mekaniskaoch termiska stabilitet, vilket har resulterat i 25 nm upplösning (halvperiod) i bilder av teststrukturer. De biologiska tillämpningarna harmöjliggjorts av en markant ökad röntgenintensitet vid provet, såväl somförbättrad driftsstabilitet. Resultaten som presenteras består, bland annat,av avbildning av kryofixerade hela celler med 10-sekundersexponeringar,avbildning av virusinfektioner i celler och kryotomografi med 20 minuterstotal exponeringtid.

Place, publisher, year, edition, pages
KTH Royal Institute of Technology, 2020. p. 72
Series
TRITA-SCI-FOU ; 2020:11
Keywords
x-ray microscopy, laboratory x-ray microscopy, water window, biological, cell imaging, virus
National Category
Physical Sciences
Research subject
Physics, Biological and Biomedical Physics; Physics, Optics and Photonics
Identifiers
urn:nbn:se:kth:diva-273716 (URN)978-91-7873-555-6 (ISBN)
Public defence
2020-08-21, FD5, Roslagstullsbacken 21, Stockholm, 12:00 (English)
Opponent
Supervisors
Available from: 2020-05-25 Created: 2020-05-25 Last updated: 2020-05-25Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMedScopus

Search in DiVA

By author/editor
Fogelqvist, EmelieKördel, MikaelÖnfelt, BjörnHertz, Hans
By organisation
Applied PhysicsBiomedical and X-ray PhysicsCellular Biophysics
In the same journal
Scientific Reports
Physical Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 285 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf