Hannover 2016 – wissenschaftliches Programm
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Q: Fachverband Quantenoptik und Photonik
Q 42: Poster: Quantum Optics and Photonics III
Q 42.48: Poster
Mittwoch, 2. März 2016, 16:30–19:00, Empore Lichthof
3D Pointillism Microscopy setup with two objectives — •Nora Schmidt1, Jana Hüve1,2, and Jürgen Klingauf1,2 — 1Institute of Medical Physics and Biophysics, Robert-Koch-Straße 31, and CeNTech, Heisenbergstraße 11, 48149 Münster — 2Authors contributed equally to this work
We have custom-built a setup for localisation microscopy techniques like PALM or STORM with two objectives. This enables us to collect two times more photons from each fluorescent molecule and therefore to increase the resolution accuracy by a factor of √2 [1]. Optional three-dimensional imaging is possible by inserting additional cylindrical lenses into the beam path.
We have characterised the localisation accuracy of this setup and found that using a second objective clearly improves our results. For 6600 photons we have obtained a localisation accuracy of 4 nm in the lateral plane and of 9 nm along the axial direction.
To further test the performance of our setup, we have imaged well-known biological structures of sub-resolution size, and have obtained results which match well with previously reported observations.
To further improve the localisation accuracy along the optical axis, we plan to use interferometric detection of the fluorescence light [2] as a second detection option. With this technique, we expect to improve the axial localisation accuracy to values similar to or even better than the localisation accuracy in the lateral plane.
[1]: K. Xu et al. Nat. Methods, 9, 185 (2012)
[2]: G. Shtengel et al. Proc. Natl. Acad. Sci. U.S.A., 106, 3125 (2009)