Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold

Abstract Fluoro-Gold is a fluorescent neuronal tracer suitable for targeted deep imaging of the nervous system. Widefield fluorescence microscopy enables visualization of Fluoro-Gold, but lacks depth discrimination. Though scanning laser confocal microscopy yields volumetric data, imaging depth is l...

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Autores principales:	Matthew Q. Miller, Iván Coto Hernández, Jenu V. Chacko, Steven Minderler, Nate Jowett
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1fc7e0ca3e394382b0ee2b08eabd655a
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spelling	oai:doaj.org-article:1fc7e0ca3e394382b0ee2b08eabd655a2021-12-02T17:41:18ZTwo-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold10.1038/s41598-021-97562-32045-2322https://doaj.org/article/1fc7e0ca3e394382b0ee2b08eabd655a2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97562-3https://doaj.org/toc/2045-2322Abstract Fluoro-Gold is a fluorescent neuronal tracer suitable for targeted deep imaging of the nervous system. Widefield fluorescence microscopy enables visualization of Fluoro-Gold, but lacks depth discrimination. Though scanning laser confocal microscopy yields volumetric data, imaging depth is limited, and optimal single-photon excitation of Fluoro-Gold requires an unconventional ultraviolet excitation line. Two-photon excitation microscopy employs ultrafast pulsed infrared lasers to image fluorophores at high-resolution at unparalleled depths in opaque tissue. Deep imaging of Fluoro-Gold-labeled neurons carries potential to advance understanding of the central and peripheral nervous systems, yet its two-photon spectral and temporal properties remain uncharacterized. Herein, we report the two-photon excitation spectrum of Fluoro-Gold between 720 and 990 nm, and its fluorescence decay rate in aqueous solution and murine brainstem tissue. We demonstrate unprecedented imaging depth of whole-mounted murine brainstem via two-photon excitation microscopy of Fluoro-Gold labeled facial motor nuclei. Optimal two-photon excitation of Fluoro-Gold within microscope tuning range occurred at 720 nm, while maximum lifetime contrast was observed at 760 nm with mean fluorescence lifetime of 1.4 ns. Whole-mount brainstem explants were readily imaged to depths in excess of 450 µm via immersion in refractive-index matching solution.Matthew Q. MillerIván Coto HernándezJenu V. ChackoSteven MinderlerNate JowettNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Matthew Q. Miller Iván Coto Hernández Jenu V. Chacko Steven Minderler Nate Jowett Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
description	Abstract Fluoro-Gold is a fluorescent neuronal tracer suitable for targeted deep imaging of the nervous system. Widefield fluorescence microscopy enables visualization of Fluoro-Gold, but lacks depth discrimination. Though scanning laser confocal microscopy yields volumetric data, imaging depth is limited, and optimal single-photon excitation of Fluoro-Gold requires an unconventional ultraviolet excitation line. Two-photon excitation microscopy employs ultrafast pulsed infrared lasers to image fluorophores at high-resolution at unparalleled depths in opaque tissue. Deep imaging of Fluoro-Gold-labeled neurons carries potential to advance understanding of the central and peripheral nervous systems, yet its two-photon spectral and temporal properties remain uncharacterized. Herein, we report the two-photon excitation spectrum of Fluoro-Gold between 720 and 990 nm, and its fluorescence decay rate in aqueous solution and murine brainstem tissue. We demonstrate unprecedented imaging depth of whole-mounted murine brainstem via two-photon excitation microscopy of Fluoro-Gold labeled facial motor nuclei. Optimal two-photon excitation of Fluoro-Gold within microscope tuning range occurred at 720 nm, while maximum lifetime contrast was observed at 760 nm with mean fluorescence lifetime of 1.4 ns. Whole-mount brainstem explants were readily imaged to depths in excess of 450 µm via immersion in refractive-index matching solution.
format	article
author	Matthew Q. Miller Iván Coto Hernández Jenu V. Chacko Steven Minderler Nate Jowett
author_facet	Matthew Q. Miller Iván Coto Hernández Jenu V. Chacko Steven Minderler Nate Jowett
author_sort	Matthew Q. Miller
title	Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
title_short	Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
title_full	Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
title_fullStr	Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
title_full_unstemmed	Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold
title_sort	two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer fluoro-gold
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1fc7e0ca3e394382b0ee2b08eabd655a
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_version_	1718379692958941184

Two-photon excitation fluorescent spectral and decay properties of retrograde neuronal tracer Fluoro-Gold

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