Two-photon excitation STED-CW microscopy

Paolo Bianchini; Benjamin Harke; Silvia Galiani; Alberto Diaspro

doi:10.1117/12.879790

3 March 2011 Two-photon excitation STED-CW microscopy

Paolo Bianchini, Benjamin Harke, Silvia Galiani, Alberto Diaspro

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 79032V (2011) https://doi.org/10.1117/12.879790
Event: SPIE BiOS, 2011, San Francisco, California, United States

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.

Citation Download 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 (3 March 2011); https://doi.org/10.1117/12.879790

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available