Eigen-analysis reveals components supporting super-resolution imaging of blinking fluorophores

This paper presents eigen-analysis of image stack of blinking fluorophores to identify the components that enable super-resolved imaging of blinking fluorophores. Eigen-analysis reveals that the contributions of spatial distribution of fluorophores and their temporal photon emission characteristics can be completely separated. While cross-emitter cross-pixel information of spatial distribution that permits super-resolution is encoded in two matrices, temporal statistics weigh the contribution of these matrices to the measured data. The properties and conditions of exploitation of these matrices are investigated. Con-temporary super-resolution imaging methods that use blinking for super-resolution are studied in the context of the presented analysis. Besides providing insight into the capabilities and limitations of existing super-resolution methods, the analysis shall help in designing better super-resolution techniques that directly exploit these matrices.

[1]  T. Lasser,et al.  Mapping molecular statistics with balanced super-resolution optical fluctuation imaging (bSOFI) , 2012, Optical Nanoscopy.

[2]  S. Volkán-Kacsó Two-state theory of binned photon statistics for a large class of waiting time distributions and its application to quantum dot blinking. , 2014, The Journal of chemical physics.

[3]  Thomas R Huser,et al.  Entropy-Based Super-Resolution Imaging (ESI): From Disorder to Fine Detail , 2015 .

[4]  R. Tsien,et al.  On/off blinking and switching behaviour of single molecules of green fluorescent protein , 1997, Nature.

[5]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[6]  Mark Bates,et al.  Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging , 2011, Nature Methods.

[7]  M. Schmid Principles Of Optics Electromagnetic Theory Of Propagation Interference And Diffraction Of Light , 2016 .

[8]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[9]  Dylan T Burnette,et al.  Bayesian localisation microscopy reveals nanoscale podosome dynamics , 2011, Nature Methods.

[10]  S. Weiss,et al.  Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI) , 2009, Proceedings of the National Academy of Sciences.

[11]  Fang Huang,et al.  Quantifying and Optimizing Single-Molecule Switching Nanoscopy at High Speeds , 2015, PloS one.

[12]  Ricardo Henriques,et al.  Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations , 2016, Nature Communications.

[13]  M. Heilemann,et al.  Carbocyanine dyes as efficient reversible single-molecule optical switch. , 2005, Journal of the American Chemical Society.

[14]  A. Efros,et al.  Random Telegraph Signal in the Photoluminescence Intensity of a Single Quantum Dot , 1997 .

[15]  Jan Vogelsang,et al.  Make them blink: probes for super-resolution microscopy. , 2010, Chemphyschem : a European journal of chemical physics and physical chemistry.

[16]  W. Webb,et al.  Precise nanometer localization analysis for individual fluorescent probes. , 2002, Biophysical journal.

[17]  A. Diaspro,et al.  Live-cell 3D super-resolution imaging in thick biological samples , 2011, Nature Methods.

[18]  Joshua W. Shaevitz,et al.  Spatial Covariance Reconstructive (SCORE) Super-Resolution Fluorescence Microscopy , 2014, PloS one.

[19]  Keith A. Lidke,et al.  Fast, single-molecule localization that achieves theoretically minimum uncertainty , 2010, Nature Methods.

[20]  S. Weiss,et al.  Achieving increased resolution and more pixels with Superresolution Optical Fluctuation Imaging (SOFI) , 2010, Optics express.

[21]  S. Holden,et al.  DAOSTORM: an algorithm for high- density super-resolution microscopy , 2011, Nature Methods.

[22]  S. Ram,et al.  Localization accuracy in single-molecule microscopy. , 2004, Biophysical journal.

[23]  Stefan Jakobs,et al.  Novel red fluorophores with superior performance in STED microscopy , 2012, Optical Nanoscopy.

[24]  Radek Macháň,et al.  Multiple signal classification algorithm for super-resolution fluorescence microscopy , 2016, Nature Communications.

[25]  Michael D. Mason,et al.  Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. , 2006, Biophysical journal.

[26]  Masaru Kuno,et al.  Universal emission intermittency in quantum dots, nanorods and nanowires , 2008, 0810.2509.

[27]  Taekjip Ha,et al.  Photophysics of fluorescent probes for single-molecule biophysics and super-resolution imaging. , 2012, Annual review of physical chemistry.

[28]  Jerker Widengren,et al.  Characterization of Photoinduced Isomerization and Back-Isomerization of the Cyanine Dye Cy5 by Fluorescence Correlation Spectroscopy , 2000 .

[29]  A. Small,et al.  Fluorophore localization algorithms for super-resolution microscopy , 2014, Nature Methods.

[30]  Sebastian van de Linde,et al.  How to switch a fluorophore: from undesired blinking to controlled photoswitching. , 2014, Chemical Society reviews.