The double-helix microscope super-resolves extended biological structures by localizing single blinking molecules in three dimensions with nanoscale precision

Hsiao Lu D. Lee, Steffen J. Sahl, Matthew D. Lew, W. E. Moerner

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

The double-helix point spread function microscope encodes the axial (z) position information of single emitters in wide-field (x,y) images, thus enabling localization in three dimensions (3D) inside extended volumes. We experimentally determine the statistical localization precision σof this approach using single emitters in a cell under typical background conditions, demonstrating σ<20 nm laterally and <30 nm axially for N ≈ 1180 photons per localization. Combined with light-induced blinking of single-molecule labels, we present proof-of-concept imaging beyond the optical diffraction limit of microtubule network structures in fixed mammalian cells over a large axial range in three dimensions.

Original languageEnglish
Article number153701
JournalApplied Physics Letters
Volume100
Issue number15
DOIs
StatePublished - Apr 9 2012

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