Site-specific 3D imaging of cells and tissues with a dual beam microscope

Jurgen A W Heymann, Mike Hayles, Ingo Gestmann, Lucille A. Giannuzzi, Ben Lich, Sriram Subramaniam

Research output: Contribution to journalArticle

Abstract

Current approaches to 3D imaging at subcellular resolution using confocal microscopy and electron tomography, while powerful, are limited to relatively thin and transparent specimens. Here we report on the use of a new generation of dual beam electron microscopes capable of site-specific imaging of the interior of cellular and tissue specimens at spatial resolutions about an order of magnitude better than those currently achieved with optical microscopy. The principle of imaging is based on using a focused ion beam to create a cut at a designated site in the specimen, followed by viewing the newly generated surface with a scanning electron beam. Iteration of these two steps several times thus results in the generation of a series of surface maps of the specimen at regularly spaced intervals, which can be converted into a three-dimensional map of the specimen. We have explored the potential of this sequential "slice-and-view" strategy for site-specific 3D imaging of frozen yeast cells and tumor tissue, and establish that this approach can identify the locations of intracellular features such as the 100 nm-wide yeast nuclear pore complex. We also show that 200 nm thick sections can be generated in situ by "milling" of resin-embedded specimens using the ion beam, providing a valuable alternative to manual sectioning of cells and tissues using an ultramicrotome. Our results demonstrate that dual beam imaging is a powerful new tool for cellular and subcellular imaging in 3D for both basic biomedical and clinical applications.

Original languageEnglish (US)
Pages (from-to)63-73
Number of pages11
JournalJournal of Structural Biology
Volume155
Issue number1
DOIs
StatePublished - Jul 2006
Externally publishedYes

Fingerprint

Yeasts
Electron Microscope Tomography
Electrons
Ions
Nuclear Pore
Confocal Microscopy
Microscopy
Neoplasms

Keywords

  • 3D cellular imaging
  • Cryo-electron microscopy
  • Dual beam imaging
  • Focused ion beam
  • Tumor tissue
  • Yeast

ASJC Scopus subject areas

  • Structural Biology

Cite this

Heymann, J. A. W., Hayles, M., Gestmann, I., Giannuzzi, L. A., Lich, B., & Subramaniam, S. (2006). Site-specific 3D imaging of cells and tissues with a dual beam microscope. Journal of Structural Biology, 155(1), 63-73. https://doi.org/10.1016/j.jsb.2006.03.006

Site-specific 3D imaging of cells and tissues with a dual beam microscope. / Heymann, Jurgen A W; Hayles, Mike; Gestmann, Ingo; Giannuzzi, Lucille A.; Lich, Ben; Subramaniam, Sriram.

In: Journal of Structural Biology, Vol. 155, No. 1, 07.2006, p. 63-73.

Research output: Contribution to journalArticle

Heymann, JAW, Hayles, M, Gestmann, I, Giannuzzi, LA, Lich, B & Subramaniam, S 2006, 'Site-specific 3D imaging of cells and tissues with a dual beam microscope', Journal of Structural Biology, vol. 155, no. 1, pp. 63-73. https://doi.org/10.1016/j.jsb.2006.03.006
Heymann JAW, Hayles M, Gestmann I, Giannuzzi LA, Lich B, Subramaniam S. Site-specific 3D imaging of cells and tissues with a dual beam microscope. Journal of Structural Biology. 2006 Jul;155(1):63-73. https://doi.org/10.1016/j.jsb.2006.03.006
Heymann, Jurgen A W ; Hayles, Mike ; Gestmann, Ingo ; Giannuzzi, Lucille A. ; Lich, Ben ; Subramaniam, Sriram. / Site-specific 3D imaging of cells and tissues with a dual beam microscope. In: Journal of Structural Biology. 2006 ; Vol. 155, No. 1. pp. 63-73.
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