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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Heintzmann, R. & Gustafsson, M. G. L. Subdiffraction resolution in continuous samples. Nature Photon. 3, 362–364 (2009).
Gustafsson, M. Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J. Microsc. 198, 82–87 (2000).
Heintzmann, R. & Cremer, C. Laterally modulated excitation microscopy: improvement of resolution by using a diffraction grating. Proc. SPIE, Optical Biopsies and Microscopic Techniques III 3568, 185–196 (1999).
Lauer, V. New approach to optical diffraction tomography yielding a vector equation of diffraction tomography and a novel tomographic microscope. J. Microsc. 205, 165–176 (2002).
Choi, W. et al. Tomographic phase microscopy. Nature Meth. 4, 717–719 (2007).
Wilson, T. Confocal Microscopy (Academic Press, 1990).
Kner, P., Chhun, B. B., Griffis, E. R., Winoto, L. & Gustafsson, M. G. L. Super-resolution video microscopy of live cells by structured illumination. Nature Meth. 6, 339–342 (2009).
Lukosz, W. Optical systems with resolving powers exceeding the classical limit. J. Opt. Soc. Am. 56, 1463–1471 (1966).
Gur, A., Fixler, D., Micó, V., Garcia, J. & Zalevsky, Z. Linear optics based nanoscopy. Opt. Express 18, 22222–22231 (2010).
Sentenac, A., Belkebir, K., Giovannini, H. & Chaumet, P. C. Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate. Opt. Lett. 33, 255–257 (2008).
Maire, G. et al. Experimental demonstration of quantitative imaging beyond Abbé's limit with optical diffraction tomography. Phys. Rev. Lett. 102, 213905 (2009).
García, J., Zalevsky, Z. & Fixler, D. Synthetic aperture superresolution by speckle pattern projection. Opt. Express 13, 6073–6078 (2005).
Sylman, D., Micó, V., García, J. & Zalevsky, Z. Random angular coding for superresolved imaging. Appl. Opt. 49, 4874–4882 (2010).
Park, Y. et al. Speckle-field digital holographic microscopy. Opt. Express 17, 12285–12292 (2009).
Sheppard, C. Super-resolution in confocal imaging. Optik 80, 53–54 (1988).
Müller, C. B. & Enderlein, J. Image scanning microscopy. Phys. Rev. Lett. 104, 198101 (2010).
Ventalon, C. & Mertz, J. Quasi-confocal fluorescence sectioning with dynamic speckle illumination. Opt. Lett. 30, 3350–3352 (2005).
Heintzmann, R., Jovin, T. M. & Cremer, C. Saturated patterned excitation microscopy—a concept for optical resolution improvement. J. Opt. Soc. Am. A 19, 1599–1609 (2002).
Gustafsson, M. Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution. Proc. Natl Acad. Sci. USA 102, 13081–13086 (2005).
Hell, S. W. & Wichmann, J. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780–782 (1994).
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).
Author information
Authors and Affiliations
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.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary information
Supplementary information (PDF 1617 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nphoton.2012.83
This article is cited by
-
Superresolution structured illumination microscopy reconstruction algorithms: a review
Light: Science & Applications (2023)
-
Transmission structured illumination microscopy with tunable frequency illumination using tilt mirror assembly
Scientific Reports (2023)
-
Calibration-free speckle matrix imaging
Light: Science & Applications (2022)
-
Hyperbolic material enhanced scattering nanoscopy for label-free super-resolution imaging
Nature Communications (2022)
-
Demystifying speckle field interference microscopy
Scientific Reports (2022)