Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure

Gleb Shtengel, James Galbraith, Catherine Galbraith, Jennifer Lippincott-Schwartz, Jennifer M. Gillette, Suliana Manley, Rachid Sougrat, Clare M. Waterman, Pakorn Kanchanawong, Michael W. Davidson, Richard D. Fetter, Harald F. Hess

Research output: Contribution to journalArticle

544 Citations (Scopus)

Abstract

Understanding molecular-scale architecture of cells requires determination of 3D locations of specific proteins with accuracy matching their nanometer-length scale. Existing electron and light microscopy techniques are limited either in molecular specificity or resolution. Here, we introduce interferometric photoactivated localization microscopy (iPALM), the combination of photoactivated localization microscopy with single-photon, simultaneous multiphase interferometry that provides sub-20-nm 3D protein localization with optimal molecular specificity. We demonstrate measurement of the 25-nm microtubule diameter, resolve the dorsal and ventral plasma membranes, and visualize the arrangement of integrin receptors within endoplasmic reticulum and adhesion complexes, 3D protein organization previously resolved only by electron microscopy. iPALM thus closes the gap between electron tomography and light microscopy, enabling both molecular specification and resolution of cellular nanoarchitecture.

Original languageEnglish (US)
Pages (from-to)3125-3130
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number9
DOIs
StatePublished - Mar 3 2009
Externally publishedYes

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Microscopy
Electron Microscopy
Electron Microscope Tomography
Interferometry
Light
Proteins
Photons
Integrins
Microtubules
Endoplasmic Reticulum
Cell Membrane

Keywords

  • Fluorescence microscopy
  • Interferometry
  • PALM
  • Photoactivated localization microscopy
  • Single molecule imaging

ASJC Scopus subject areas

  • General

Cite this

Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure. / Shtengel, Gleb; Galbraith, James; Galbraith, Catherine; Lippincott-Schwartz, Jennifer; Gillette, Jennifer M.; Manley, Suliana; Sougrat, Rachid; Waterman, Clare M.; Kanchanawong, Pakorn; Davidson, Michael W.; Fetter, Richard D.; Hess, Harald F.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, No. 9, 03.03.2009, p. 3125-3130.

Research output: Contribution to journalArticle

Shtengel, G, Galbraith, J, Galbraith, C, Lippincott-Schwartz, J, Gillette, JM, Manley, S, Sougrat, R, Waterman, CM, Kanchanawong, P, Davidson, MW, Fetter, RD & Hess, HF 2009, 'Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure', Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 9, pp. 3125-3130. https://doi.org/10.1073/pnas.0813131106
Shtengel, Gleb ; Galbraith, James ; Galbraith, Catherine ; Lippincott-Schwartz, Jennifer ; Gillette, Jennifer M. ; Manley, Suliana ; Sougrat, Rachid ; Waterman, Clare M. ; Kanchanawong, Pakorn ; Davidson, Michael W. ; Fetter, Richard D. ; Hess, Harald F. / Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure. In: Proceedings of the National Academy of Sciences of the United States of America. 2009 ; Vol. 106, No. 9. pp. 3125-3130.
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