OperaV1, PubMed, Curation, bibRecord, 000021

Two-Photon Excitation STED Microscopy with Time-Gated Detection.

Identifieur interne : 000021 ( PubMed/Curation ); précédent : 000020; suivant : 000022

Two-Photon Excitation STED Microscopy with Time-Gated Detection.

Auteurs : Iván Coto Hernández [Italie] ; Marco Castello [Italie] ; Luca Lanzan [Italie] ; Marta D'Amora [Italie] ; Paolo Bianchini [Italie] ; Alberto Diaspro [Italie] ; Giuseppe Vicidomini [Italie]

Source :

Scientific reports ; 2016.

RBID : pubmed:26757892

Abstract

We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave. The continuous-wave stimulated emission beam tempers the laser architecture's complexity and cost, but the time-gated detection degrades the signal-to-noise ratio (SNR) and signal-to-background ratio (SBR) of the image. We recover the SNR and the SBR through a multi-image deconvolution algorithm. Indeed, the algorithm simultaneously reassigns early-photons (normally discarded by the time-gated detection) to their original positions and removes the background induced by the stimulated emission beam. We exemplify the benefits of this implementation by imaging sub-cellular structures. Finally, we discuss of the extension of this algorithm to future all-pulsed 2PE-STED implementationd based on time-gated detection and a nanosecond laser source.

DOI: 10.1038/srep19419
PubMed: 26757892

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<front><div type="abstract" xml:lang="en">We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave. The continuous-wave stimulated emission beam tempers the laser architecture's complexity and cost, but the time-gated detection degrades the signal-to-noise ratio (SNR) and signal-to-background ratio (SBR) of the image. We recover the SNR and the SBR through a multi-image deconvolution algorithm. Indeed, the algorithm simultaneously reassigns early-photons (normally discarded by the time-gated detection) to their original positions and removes the background induced by the stimulated emission beam. We exemplify the benefits of this implementation by imaging sub-cellular structures. Finally, we discuss of the extension of this algorithm to future all-pulsed 2PE-STED implementationd based on time-gated detection and a nanosecond laser source.</div>
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<Abstract><AbstractText>We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave. The continuous-wave stimulated emission beam tempers the laser architecture's complexity and cost, but the time-gated detection degrades the signal-to-noise ratio (SNR) and signal-to-background ratio (SBR) of the image. We recover the SNR and the SBR through a multi-image deconvolution algorithm. Indeed, the algorithm simultaneously reassigns early-photons (normally discarded by the time-gated detection) to their original positions and removes the background induced by the stimulated emission beam. We exemplify the benefits of this implementation by imaging sub-cellular structures. Finally, we discuss of the extension of this algorithm to future all-pulsed 2PE-STED implementationd based on time-gated detection and a nanosecond laser source.</AbstractText>
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<AffiliationInfo><Affiliation>Department of Informatics, Bioengineering, Robotics and Systems Engineering, University of Genoa, Via Opera Pia 13, 16145, Genoa, Italy.</Affiliation>
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<AffiliationInfo><Affiliation>Nikon Imaging Center, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy.</Affiliation>
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Serveur d'exploration sur l'opéra

Two-Photon Excitation STED Microscopy with Time-Gated Detection.

Two-Photon Excitation STED Microscopy with Time-Gated Detection.

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