Nonlinear structured illumination microscopy by surface plasmon enhanced stimulated emission depletion.

Nonlinear structured illumination microscopy (SIM) in theory has unlimited resolution over a full field of view. However under a realistic signal-to-noise ratio and a limited photon budget, the performance of nonlinear SIM strongly depends on the behavior of the nonlinear effect. Saturated SIM (SSIM) is not ideal in biological applications due to its strong photobleaching. Stimulated emission depletion (STED) SIM will have high sensitivity, higher resolution and less photo toxicity than SSIM. However, the laser power necessary to support a strong full-field STED effect is not attainable with current laser technology. We experimentally proved that surface plasmon resonance enhances (SPR) near surface STED effect by a factor of 8, and therefore STED-SIM is feasible in the total internal reflection microscopy mode with SPR enhancement. Simulation analysis predicts that SPR enhanced 2D STED is strong enough for nonlinear SIM to achieve high-speed imaging at 30-nm resolution and single molecule sensitivity. The STED-SIM superresolution microscopy method would provide a solution for observing single molecule processes in vitro or on the basal membrane of live cells.

[1]  S. Hell,et al.  Fluorescence nanoscopy by ground-state depletion and single-molecule return , 2008, Nature Methods.

[2]  M. Gustafsson,et al.  Subdiffraction resolution in continuous samples , 2009 .

[3]  Christian Eggeling,et al.  Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Andreas Schönle,et al.  Resolution scaling in STED microscopy. , 2008, Optics express.

[5]  C. Kuang,et al.  Far-field Optical Nanoscopy via Visible Light , 2013 .

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

[7]  S. Hell,et al.  STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis , 2006, Nature.

[8]  Mark Bates,et al.  Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy , 2008, Science.

[9]  Christian Eggeling,et al.  STED microscopy reveals crystal colour centres with nanometric resolution. , 2009 .

[10]  S. Hell,et al.  Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[11]  R. Heintzmann,et al.  Saturated patterned excitation microscopy--a concept for optical resolution improvement. , 2002, Journal of the Optical Society of America. A, Optics, image science, and vision.

[12]  X. Zhuang,et al.  Breaking the Diffraction Barrier: Super-Resolution Imaging of Cells , 2010, Cell.

[13]  Shean-Jen Chen,et al.  Surface plasmon-enhanced two-photon fluorescence microscopy for live cell membrane imaging. , 2009 .

[14]  T. Bonhoeffer,et al.  Live-cell imaging of dendritic spines by STED microscopy , 2008, Proceedings of the National Academy of Sciences.

[15]  Y.-D. Su,et al.  Surface plasmon-enhanced two-photon fluorescence microscopy for live cell membrane imaging , 2009, BiOS.

[16]  S. Hell,et al.  STED microscopy with continuous wave beams , 2007, Nature Methods.

[17]  S. Hell.  Microscopy and its focal switch , 2008, Nature Methods.

[18]  R. Dasari,et al.  Single Molecule Detection Using Surface-Enhanced Raman Scattering (SERS) , 1997 .

[19]  Stefan W. Hell,et al.  Single-molecule STED microscopy with photostable organic fluorophores. , 2010, Small.

[20]  Colin J R Sheppard,et al.  Wide-field extended-resolution fluorescence microscopy with standing surface-plasmon-resonance waves. , 2009, Optics letters.

[21]  Stefan W. Hell,et al.  Supporting Online Material Materials and Methods Figs. S1 to S9 Tables S1 and S2 References Video-rate Far-field Optical Nanoscopy Dissects Synaptic Vesicle Movement , 2022 .

[22]  S. Hell,et al.  Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. , 1994, Optics letters.

[23]  M. Gustafsson.  Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy , 2000, Journal of microscopy.

[24]  Liisa M. Hirvonen,et al.  Structured illumination microscopy using photoswitchable fluorescent proteins , 2008, SPIE BiOS.

[25]  Vladimir M. Shalaev,et al.  Searching for better plasmonic materials , 2009, 0911.2737.

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

[27]  Chi-Hung Lin,et al.  Enhanced live cell membrane imaging using surface plasmon-enhanced total internal reflection fluorescence microscopy. , 2006, Optics express.

[28]  Wolfgang Knoll,et al.  Surface-Plasmon Field-Enhanced Fluorescence Spectroscopy , 2000 .

[29]  P. So,et al.  Resolution enhancement in standing-wave total internal reflection microscopy: a point-spread-function engineering approach. , 2001, Journal of the Optical Society of America. A, Optics, image science, and vision.

[30]  W. Knoll,et al.  Fluorescence intensity in surface-plasmon field-enhanced fluorescence spectroscopy , 2005 .

[31]  S. Hell,et al.  Stimulated emission depletion (STED) nanoscopy of a fluorescent protein-labeled organelle inside a living cell , 2008, Proceedings of the National Academy of Sciences.

[32]  M. Gustafsson.  Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Bryant B. Chhun,et al.  Super-Resolution Video Microscopy of Live Cells by Structured Illumination , 2009, Nature Methods.

[34]  Jiří Homola,et al.  Electromagnetic Theory of Surface Plasmons , 2006 .

[35]  Herbert Schneckenburger,et al.  Light Dose is a Limiting Factor to Maintain Cell Viability in Fluorescence Microscopy and Single Molecule Detection , 2010, International journal of molecular sciences.

[36]  Rainer Heintzmann,et al.  Saturated patterned excitation microscopy with two-dimensional excitation patterns. , 2003, Micron.

[37]  Mark Bates,et al.  Super-resolution fluorescence microscopy. , 2009, Annual review of biochemistry.