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Proceedings Paper

The importance of the photon arrival times in STED microscopy
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Paper Abstract

In a stimulated emission depletion (STED) microscope the region from which a fluorophore can spontaneously emit shrinks with the continued STED beam action after the excitation event. This fact has been recently used to implement a versatile, simple and cheap STED microscope that uses a pulsed excitation beam, a STED beam running in continuous-wave (CW) and a time-gated detection: By collecting only the delayed (with respect to the excitation events) fluorescence, the STED beam intensity needed for obtaining a certain spatial resolution strongly reduces, which is fundamental to increase live cell imaging compatibility. This new STED microscopy implementation, namely gated CW-STED, is in essence limited (only) by the reduction of the signal associated with the time-gated detection. Here we show the recent advances in gated CW-STED microscopy and related methods. We show that the time-gated detection can be substituted by more efficient computational methods when the arrival-times of all fluorescence photons are provided.

Paper Details

Date Published: 9 March 2015
PDF: 7 pages
Proc. SPIE 9331, Single Molecule Spectroscopy and Superresolution Imaging VIII, 93310X (9 March 2015); doi: 10.1117/12.2081553
Show Author Affiliations
Marco Castello, Istituto Italiano di Tecnologia (Italy)
DIBRIS, Univ. of Genoa (Italy)
Luca L. Lanzanò, Istituto Italiano di Tecnologia (Italy)
Iván Coto Hernández, Istituto Italiano di Tecnologia (Italy)
DIFI, Univ. of Genoa (Italy)
Christian Eggeling, Univ. of Oxford (United Kingdom)
Alberto Diaspro, Istituto Italiano di Tecnologia (Italy)
DIFI, Univ. of Genoa (Italy)
Nikon Imaging Ctr. (Italy)
Giuseppe Vicidomini, Istituto Italiano di Tecnologia (Italy)

Published in SPIE Proceedings Vol. 9331:
Single Molecule Spectroscopy and Superresolution Imaging VIII
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)

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