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Proceedings Article

Two-photon excitation STED-CW microscopy

[+] Author Affiliations
Paolo Bianchini, Silvia Galiani, Alberto Diaspro

Italian Institute of Technology (Italy) and Univ. of Genoa (Italy)

Benjamin Harke

Italian Institute of Technology (Italy)

Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032V (March 03, 2011); doi:10.1117/12.879790
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From Conference Volume 7903

  • Multiphoton Microscopy in the Biomedical Sciences XI
  • Ammasi Periasamy; Karsten König; Peter T. C. So
  • San Francisco, California, USA | January 22, 2011

abstract

Here, we report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with continuous-wave (CW) stimulated-emission depletion (STED). We show an easy-to- implement and promising laser combination based on Ti:Sapphire ultrafast laser source for two-photon excitation and a commercial Leica TCS STED-CW microscope for resolution enhancement. Images of fluorescent nanoparticles produce comparative similar resolution to the one photon excitation. Two-photon excitation STED microscopy achieves approximately 3-4 fold improvement in resolution in the radial direction over conventional 2PELSM. Further improvements in resolution are theoretically achievable, suggesting that 2PE STED microscopy will permit nanoscale imaging, for instance, of neuronal structures located in relatively intact brain tissue.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Paolo Bianchini ; Benjamin Harke ; Silvia Galiani and Alberto Diaspro
"Two-photon excitation STED-CW microscopy", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032V (March 03, 2011); doi:10.1117/12.879790; http://dx.doi.org/10.1117/12.879790


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