TY - JOUR
T1 - Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure
AU - Shtengel, Gleb
AU - Galbraith, James A.
AU - Galbraith, Catherine G.
AU - Lippincott-Schwartz, Jennifer
AU - Gillette, Jennifer M.
AU - Manley, Suliana
AU - Sougrat, Rachid
AU - Waterman, Clare M.
AU - Kanchanawong, Pakorn
AU - Davidson, Michael W.
AU - Fetter, Richard D.
AU - Hess, Harald F.
PY - 2009/3/3
Y1 - 2009/3/3
N2 - 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.
AB - 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.
KW - Fluorescence microscopy
KW - Interferometry
KW - PALM
KW - Photoactivated localization microscopy
KW - Single molecule imaging
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U2 - 10.1073/pnas.0813131106
DO - 10.1073/pnas.0813131106
M3 - Article
C2 - 19202073
AN - SCOPUS:62549155396
SN - 0027-8424
VL - 106
SP - 3125
EP - 3130
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 9
ER -