Aberration Characterization and Correction in Super-Resolution Localization Microscopy
(2.Key Laboratory for Biomedical Photonics, Department of Biomedical Engineering, Huazhong University of Science and Technology, Wuhan, Hubei, China 430074)
【Abstract】Super-resolution localization microscopy can achieve the ultra-high spatial resolution up to several nanometers by the single molecule localization and reconstruction from thousands or even tens of thousands of single molecule image, which provides unprecedented opportunities for studying the cell structures and biological phenomenon. However, the aberration (originating from the imperfection of the optical system or the inhomogeneity of the sample itself) distorts the raw images from single molecules, which decreases the final spatial resolution and even results in wrong results. The effects of several representative aberrations on super-resolution localization imaging are quantitatively characterized, and an aberration correction method based on the sample image itself is proposed. Simulation and experimental results show that the aberrations cause distortion of the point spread function and the decrease of the spatial resolution. The image quality can be restored by using the proposed aberration correction method.
【Keywords】 microscopy; fluorescence microscopy; super-resolution imaging; aberration correction; resolution; localization precision;
Betzig E, Patterson G H, Sougrat R, et al. Imaging intracellular fluorescent proteins at nanometer resolution[J]. Science, 2006, 313 (5793): 1642-1645.
Hess S T, Girirajan T P K, Mason M D. Ultra-high resolution imaging by fluorescence photoactivation localization microscopy[J]. Biophysical Journal, 2006, 91 (11): 4258-4272.
Rust M J, Bates M, Zhuang X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)[J]. Nature Methods, 2006, 3(10): 793-795.
Hell S W, Wichmann J. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy[J]. Optics Letters, 1994, 19 (11): 780-782.
Gustafsson M G L. Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102 (37): 13081-13086.
Huang B, Jones S A, Brandenburg B, et al. Whole-cell 3DSTORM reveals interactions between cellular structures with nanometer-scale resolution[J]. Nature Methods, 2008, 5 (12): 1047-1052.
Schwertner M, Booth M J, Wilson T. Characterizing specimen induced aberrations for high NA adaptive optical microscopy[J]. Optics Express, 2004, 12 (26): 6540-6552.
Hell S, Reiner G, Cremer C, et al. Aberrations in confocal fluorescence microscopy induced by mismatches in refractive index[J]. Journal of Microscopy, 1993, 169 (3): 391-405.
Quirin S, Pavani S R P, Piestun R. Optimal 3D single-molecule localization for superresolution microscopy with aberrations and engineered point spread functions[J]. Proceedings of the National Academy of Sciences, 2012, 109 (3): 675-679.
McGorty R, Schnitzbauer J, Zhang W, et al. Correction of depth-dependent aberrations in 3D single-molecule localization and super-resolution microscopy[J]. Optics Letters, 2014, 39 (2): 275-278.
Burke D, Patton B, Huang F, et al. Adaptive optics correction of specimen-induced aberrations in single-molecule switching microscopy[J]. Optica, 2015, 2(2): 177-185.
Tehrani K F, Xu J, Zhang Y, et al. Adaptive optics stochastic optical reconstruction microscopy (AO-STORM) using agenetic algorithm[J]. Optics Express, 2015, 23(10): 13677-13692.
Booth M, Andrade D, Burke D, et al. Aberrations and adaptive optics in super-resolution microscopy[J]. Microscopy (Oxford), 2015, 64 (4): 251-261.
Noll R J. Zernike polynomials and atmospheric turbulence[J]. Journal of the Optical Society of America, 1976, 66 (3): 207-211.
Coles B C, Webb S E D, Schwartz N, et al. Characterisation of the effects of optical aberrations in single molecule techniques[J]. Biomedical Optics Express, 2016, 7 (5): 1755-1767.
Biomedical Imaging Group, Ecole Polytechnique Fédérale de Lausanne. Single-molecule localization microscopy·software benchmarking[EB/OL]. (2016-08-19)[2016-09-29]. http: //bigwww. epfl. ch/palm/datasets/index. html?p=usage.
Nieuwenhuizen R P, Lidke K A, Bates M, et al. Measuring image resolution in optical nanoscopy[J]. Nature Methods2013, 10 (6): 557-562.