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Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy

In recent years, fluorescent nanomaterials have gained high relevance in biological applications as probes for various fluorescence-based spectroscopy and imaging techniques. Among these materials, dye-doped silica nanoparticles have demonstrated a high potential to overcome the limitations presente...

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Autores principales: Tavernaro, Isabella, Cavelius, Christian, Peuschel, Henrike, Kraegeloh, Annette
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Beilstein-Institut 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496580/
https://www.ncbi.nlm.nih.gov/pubmed/28690964
http://dx.doi.org/10.3762/bjnano.8.130
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author Tavernaro, Isabella
Cavelius, Christian
Peuschel, Henrike
Kraegeloh, Annette
author_facet Tavernaro, Isabella
Cavelius, Christian
Peuschel, Henrike
Kraegeloh, Annette
author_sort Tavernaro, Isabella
collection PubMed
description In recent years, fluorescent nanomaterials have gained high relevance in biological applications as probes for various fluorescence-based spectroscopy and imaging techniques. Among these materials, dye-doped silica nanoparticles have demonstrated a high potential to overcome the limitations presented by conventional organic dyes such as high photobleaching, low stability and limited fluorescence intensity. In the present work we describe an effective approach for the preparation of fluorescent silica nanoparticles in the size range between 15 and 80 nm based on L-arginine-controlled hydrolysis of tetraethoxysilane in a biphasic cyclohexane–water system. Commercially available far-red fluorescent dyes (Atto647N, Abberior STAR 635, Dy-647, Dy-648 and Dy-649) were embedded covalently into the particle matrix, which was achieved by aminosilane coupling. The physical particle attributes (particle size, dispersion, degree of agglomeration and stability) and the fluorescence properties of the obtained particles were compared to particles from commonly known synthesis methods. As a result, the spectroscopic characteristics of the presented monodisperse dye-doped silica nanoparticles were similar to those of the free uncoupled dyes, but indicate a much higher photostability and brightness. As revealed by dynamic light scattering and ζ-potential measurements, all particle suspensions were stable in water and cell culture medium. In addition, uptake studies on A549 cells were performed, using confocal and stimulated emission depletion (STED) microscopy. Our approach allows for a step-by-step formation of dye-doped silica nanoparticles in the form of dye-incorporated spheres, which can be used as versatile fluorescent probes in confocal and STED imaging.
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spelling pubmed-54965802017-07-07 Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy Tavernaro, Isabella Cavelius, Christian Peuschel, Henrike Kraegeloh, Annette Beilstein J Nanotechnol Full Research Paper In recent years, fluorescent nanomaterials have gained high relevance in biological applications as probes for various fluorescence-based spectroscopy and imaging techniques. Among these materials, dye-doped silica nanoparticles have demonstrated a high potential to overcome the limitations presented by conventional organic dyes such as high photobleaching, low stability and limited fluorescence intensity. In the present work we describe an effective approach for the preparation of fluorescent silica nanoparticles in the size range between 15 and 80 nm based on L-arginine-controlled hydrolysis of tetraethoxysilane in a biphasic cyclohexane–water system. Commercially available far-red fluorescent dyes (Atto647N, Abberior STAR 635, Dy-647, Dy-648 and Dy-649) were embedded covalently into the particle matrix, which was achieved by aminosilane coupling. The physical particle attributes (particle size, dispersion, degree of agglomeration and stability) and the fluorescence properties of the obtained particles were compared to particles from commonly known synthesis methods. As a result, the spectroscopic characteristics of the presented monodisperse dye-doped silica nanoparticles were similar to those of the free uncoupled dyes, but indicate a much higher photostability and brightness. As revealed by dynamic light scattering and ζ-potential measurements, all particle suspensions were stable in water and cell culture medium. In addition, uptake studies on A549 cells were performed, using confocal and stimulated emission depletion (STED) microscopy. Our approach allows for a step-by-step formation of dye-doped silica nanoparticles in the form of dye-incorporated spheres, which can be used as versatile fluorescent probes in confocal and STED imaging. Beilstein-Institut 2017-06-21 /pmc/articles/PMC5496580/ /pubmed/28690964 http://dx.doi.org/10.3762/bjnano.8.130 Text en Copyright © 2017, Tavernaro et al. https://creativecommons.org/licenses/by/4.0https://www.beilstein-journals.org/bjnano/termsThis is an Open Access article under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The license is subject to the Beilstein Journal of Nanotechnology terms and conditions: (https://www.beilstein-journals.org/bjnano/terms)
spellingShingle Full Research Paper
Tavernaro, Isabella
Cavelius, Christian
Peuschel, Henrike
Kraegeloh, Annette
Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title_full Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title_fullStr Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title_full_unstemmed Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title_short Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
title_sort bright fluorescent silica-nanoparticle probes for high-resolution sted and confocal microscopy
topic Full Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496580/
https://www.ncbi.nlm.nih.gov/pubmed/28690964
http://dx.doi.org/10.3762/bjnano.8.130
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