Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.

Continuous improvements in imaging techniques are challenging biologists to search for more accurate methods to label cellular elements. This is particularly relevant for diffraction-unlimited fluorescence imaging, where the perceived resolution is affected by the size of the affinity probes. This i...

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Main Authors: Maria Angela Gomes de Castro, Claudia Höbartner, Felipe Opazo
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC5325610?pdf=render
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spelling doaj-8f2aeb2459014e6ba0431647de6b85752020-11-24T20:41:39ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01122e017305010.1371/journal.pone.0173050Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.Maria Angela Gomes de CastroClaudia HöbartnerFelipe OpazoContinuous improvements in imaging techniques are challenging biologists to search for more accurate methods to label cellular elements. This is particularly relevant for diffraction-unlimited fluorescence imaging, where the perceived resolution is affected by the size of the affinity probes. This is evident when antibodies, which are 10-15 nm in size, are used. Previously it has been suggested that RNA aptamers (~3 nm) can be used to detect cellular proteins under super-resolution imaging. However, a direct comparison between several aptamers and antibodies is needed, to clearly show the advantages and/or disadvantages of the different probes. Here we have conducted such a comparative study, by testing several aptamers and antibodies using stimulated emission depletion microscopy (STED). We have targeted three membrane receptors, EGFR, ErbB2 and Epha2, which are relevant to human health, and recycle between plasma membrane and intracellular organelles. Our results suggest that the aptamers can reveal more epitopes than most antibodies, thus providing a denser labeling of the stained structures. Moreover, this improves the overall quality of the information that can be extracted from the images. We conclude that aptamers could become useful fluorescent labeling tools for light microscopy and super-resolution imaging, and that their development for novel targets is imperative.http://europepmc.org/articles/PMC5325610?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Maria Angela Gomes de Castro
Claudia Höbartner
Felipe Opazo
spellingShingle Maria Angela Gomes de Castro
Claudia Höbartner
Felipe Opazo
Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
PLoS ONE
author_facet Maria Angela Gomes de Castro
Claudia Höbartner
Felipe Opazo
author_sort Maria Angela Gomes de Castro
title Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
title_short Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
title_full Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
title_fullStr Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
title_full_unstemmed Aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
title_sort aptamers provide superior stainings of cellular receptors studied under super-resolution microscopy.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2017-01-01
description Continuous improvements in imaging techniques are challenging biologists to search for more accurate methods to label cellular elements. This is particularly relevant for diffraction-unlimited fluorescence imaging, where the perceived resolution is affected by the size of the affinity probes. This is evident when antibodies, which are 10-15 nm in size, are used. Previously it has been suggested that RNA aptamers (~3 nm) can be used to detect cellular proteins under super-resolution imaging. However, a direct comparison between several aptamers and antibodies is needed, to clearly show the advantages and/or disadvantages of the different probes. Here we have conducted such a comparative study, by testing several aptamers and antibodies using stimulated emission depletion microscopy (STED). We have targeted three membrane receptors, EGFR, ErbB2 and Epha2, which are relevant to human health, and recycle between plasma membrane and intracellular organelles. Our results suggest that the aptamers can reveal more epitopes than most antibodies, thus providing a denser labeling of the stained structures. Moreover, this improves the overall quality of the information that can be extracted from the images. We conclude that aptamers could become useful fluorescent labeling tools for light microscopy and super-resolution imaging, and that their development for novel targets is imperative.
url http://europepmc.org/articles/PMC5325610?pdf=render
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AT claudiahobartner aptamersprovidesuperiorstainingsofcellularreceptorsstudiedundersuperresolutionmicroscopy
AT felipeopazo aptamersprovidesuperiorstainingsofcellularreceptorsstudiedundersuperresolutionmicroscopy
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