4Pi-SMS microscopy

In the simplest case, an SMS image is a histogram of sufficiently many marker positions, each being calculated from the diffraction-limited fluorescence image of a single emitter. Here, the localization in the dimension along the optic axis poses a challenge. The 4Pi-SMS microscope solves this problem by using two opposing objective lenses coherently for fluorescence detection [Aquino et al., 2011]. The analysis of at least three interference patterns for each marker, each exhibiting a different relative phase, allows precise axial localization.

The 4Pi-SMS microscope can localize markers with <10 nm precision in three dimensions in an extended layer of 650 nm thickness. It can also distinguish between different species of markers, e.g. markers of different color.

 

Further information:

  • D. Aquino, A. Schönle, C. Geisler, C. v. Middendorff, C. A. Wurm, Y. Okamura, T. Lang, S. W. Hell, A. Egner:
    "Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores", Nature Methods, 8 (4), 353 - 359, 2009
  • S. W. Hell, R. Schmidt, A. Egner:
    "Diffraction-unlimited three-dimensional optical nanoscopy with opposing lenses",  Nature Photonics 3, 381 - 387, 2009
  • C. v. Middendorff, A. Egner, C. Geisler, S. W. Hell, A. Schönle:
    "Isotropic 3D Nanoscopy based on single emitter switching"
    Opt. Expr. 16 (25), 20774-20788, 2008

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Laser-Laboratorium Göttingen e.V. (LLG)

Contact person:

Head of department
Prof. Dr. Alexander Egner
"Optical Nanoscopy"

Tel.: +49(0)551/5035-35
FAX: +49(0)551/5035-99
alexander.egner(at)llg-ev.de