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Structured illumination microscopy using unknown speckle patterns

Nature Photonics volume 6, pages 312–315 (2012)Cite this article

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Abstract

Using spatially non-uniform illumination significantly improves the resolution of light microscopy1. Indeed, frequency mixing between the object and the illumination permits the recovery of object frequencies beyond the diffraction-limited detection band pass2,3,4,5. However, the image reconstruction process requires a precise knowledge of the illumination patterns (usually focused or periodic) and therefore sophisticated stable mountings6,7. Here, we show, both theoretically and experimentally, that image reconstruction can be performed without knowing the illumination patterns, provided that their average is roughly homogeneous over the sample. Using blind structured illumination microscopy (blind-SIM), a resolution about two times better than that of conventional wide-field microscopy is obtained by simply illuminating the sample with several uncontrolled random speckles. Our approach is insensitive to specimen or aberration-induced illumination deformations, does not require any calibration step or stringent control of the illumination, and dramatically simplifies the experimental set-up.

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Figure 1: Illustration of the different steps of a simulated speckle blind-SIM experiment.
Figure 2: Numerical study of the resolution of the reconstructed sample using speckle blind-SIM versus the excitation pattern spectrum.
Figure 3: Experimental periodical SIM data processed by blind-SIM.
Figure 4: Experimental speckle data processed by blind-SIM.

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Acknowledgements

The authors thank R. Heintzmann for a very nice suggestion concerning the blind-SIM algorithm. The authors also thank the anonymous referees for their constructive and stimulating remarks. This work was funded by the French Agence National de la Recherche (contract no. ANR-08-NANO-P053-36).

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Authors and Affiliations

  1. Institut Fresnel (CNRS UMR 7249), Université Aix-Marseille, Campus de St Jérôme, Marseille, 13013, France

    E. Mudry, K. Belkebir, J. Girard, J. Savatier, E. Le Moal, M. Allain & A. Sentenac

  2. ISM2 (CNRS UMR 7313) Université Aix-Marseille, Campus de St Jérôme, Marseille, 13013, France

    C. Nicoletti

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  1. E. Mudry
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  8. A. Sentenac
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Contributions

E.M., K.B., J.G. and A.S. conceived the blind-SIM approach and wrote the manuscript. E.M. and K.B. wrote the code and ran the model. J.G. performed the experiments and pre-processed the data. J.S. and C.N. prepared the samples. E.Le.M. and M.A. provided technical support and A.S. supervised the project.

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Correspondence to A. Sentenac.

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The authors declare no competing financial interests.

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Mudry, E., Belkebir, K., Girard, J. et al. Structured illumination microscopy using unknown speckle patterns. Nature Photon 6, 312–315 (2012). https://doi.org/10.1038/nphoton.2012.83

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