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Proceedings Paper

Two-photon excitation STED-CW microscopy
Author(s): Paolo Bianchini; Benjamin Harke; Silvia Galiani; Alberto Diaspro
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Paper 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.

Paper Details

Date Published: 3 March 2011
PDF: 5 pages
Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032V (3 March 2011); doi: 10.1117/12.879790
Show Author Affiliations
Paolo Bianchini, Italian Institute of Technology (Italy)
Univ. of Genoa (Italy)
Benjamin Harke, Italian Institute of Technology (Italy)
Silvia Galiani, Italian Institute of Technology (Italy)
Univ. of Genoa (Italy)
Alberto Diaspro, Italian Institute of Technology (Italy)
Univ. of Genoa (Italy)

Published in SPIE Proceedings Vol. 7903:
Multiphoton Microscopy in the Biomedical Sciences XI
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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