Accurate axial localization by conical diffraction beam shaping generating a dark-helix PSF

Clement Fallet; Astrid Lassalle; Maxime Dubois-Delumeau; Gabriel Y. Sirat

doi:10.1117/12.2212215

1 March 2016 Accurate axial localization by conical diffraction beam shaping generating a dark-helix PSF

Clement Fallet, Astrid Lassalle, Maxime Dubois-Delumeau, Gabriel Y. Sirat

Proceedings Volume 9714, Single Molecule Spectroscopy and Superresolution Imaging IX; 971414 (2016) https://doi.org/10.1117/12.2212215
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

We present here a new PSF-shaping technique using biaxial crystals to generate a highly z-dependent distribution in single molecule localization microscopy (SMLM). This distribution features two zeros of intensity that rotate together with defocus. This PSF features similarities to the double-helix introduced by Moerner and Piestun and thus has been dubbed dark-helix since we track zeros of intensity. Preliminary numerical studies based on Cramer-Rao Lower Bound (CRLB) show that this PSF has the potential to obtain up to 20nm localization precision. This PSF can be easily generated by a very simple, monolithic add-on added in front of the detection camera. Additionally, the PSF remains of the approximate size of the Airy PSF, the x-y localization precision is not substantially affected and no trade-off is required. The xy compacity of the PSF also enables theoretically a higher density of emitters than the double-helix which spreads on a larger scale. Limiting factors for SMLM such as loss of photons, complexity and robustness will be discussed and considerations about the practical implementation of such techniques will be given.

Citation Download Citation

Clement Fallet, Astrid Lassalle, Maxime Dubois-Delumeau, and Gabriel Y. Sirat "Accurate axial localization by conical diffraction beam shaping generating a dark-helix PSF", Proc. SPIE 9714, Single Molecule Spectroscopy and Superresolution Imaging IX, 971414 (1 March 2016); https://doi.org/10.1117/12.2212215

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