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Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence
The dynamics of living cells can be studied by live-cell fluorescence microscopy. However, this requires the use of excessive light energy to obtain good signal-to-noise ratio, which can then photobleach fluorochromes, and more worrisomely, lead to phototoxicity. Upon light excitation, noble metal n...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10551705/ https://www.ncbi.nlm.nih.gov/pubmed/37405751 http://dx.doi.org/10.1091/mbc.E22-06-0200 |
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author | Soha, Sumaiya A. Santhireswaran, Araniy Huq, Saaimatul Casimir-Powell, Jayde Jenkins, Nicala Hodgson, Gregory K. Sugiyama, Michael Antonescu, Costin N. Impellizzeri, Stefania Botelho, Roberto J. |
author_facet | Soha, Sumaiya A. Santhireswaran, Araniy Huq, Saaimatul Casimir-Powell, Jayde Jenkins, Nicala Hodgson, Gregory K. Sugiyama, Michael Antonescu, Costin N. Impellizzeri, Stefania Botelho, Roberto J. |
author_sort | Soha, Sumaiya A. |
collection | PubMed |
description | The dynamics of living cells can be studied by live-cell fluorescence microscopy. However, this requires the use of excessive light energy to obtain good signal-to-noise ratio, which can then photobleach fluorochromes, and more worrisomely, lead to phototoxicity. Upon light excitation, noble metal nanoparticles such as silver nanoparticles (AgNPs) generate plasmons, which can then amplify excitation in direct proximity of the nanoparticle’s surface and couple to the oscillating dipole of nearby radiating fluorophores, modifying their rate of emission and thus, enhancing their fluorescence. Here, we show that AgNPs fed to cells to accumulate within lysosomes enhanced the fluorescence of lysosome-targeted Alexa488-conjugated dextran, BODIPY-cholesterol, and DQ-BSA. Moreover, AgNP increased the fluorescence of GFP fused to the cytosolic tail of LAMP1, showing that metal enhanced fluorescence can occur across the lysosomal membrane. The inclusion of AgNPs in lysosomes did not disturb lysosomal properties such as lysosomal pH, degradative capacity, autophagy and autophagic flux, and membrane integrity, though AgNP seemed to increase basal lysosome tubulation. Importantly, by using AgNP, we could track lysosome motility with reduced laser power without damaging and altering lysosome dynamics. Overall, AgNP-enhanced fluorescence may be a useful tool to study the dynamics of the endo-lysosomal pathway while minimizing phototoxicity. |
format | Online Article Text |
id | pubmed-10551705 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-105517052023-11-01 Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence Soha, Sumaiya A. Santhireswaran, Araniy Huq, Saaimatul Casimir-Powell, Jayde Jenkins, Nicala Hodgson, Gregory K. Sugiyama, Michael Antonescu, Costin N. Impellizzeri, Stefania Botelho, Roberto J. Mol Biol Cell Special Issue on Cell Biology of the Lysosome The dynamics of living cells can be studied by live-cell fluorescence microscopy. However, this requires the use of excessive light energy to obtain good signal-to-noise ratio, which can then photobleach fluorochromes, and more worrisomely, lead to phototoxicity. Upon light excitation, noble metal nanoparticles such as silver nanoparticles (AgNPs) generate plasmons, which can then amplify excitation in direct proximity of the nanoparticle’s surface and couple to the oscillating dipole of nearby radiating fluorophores, modifying their rate of emission and thus, enhancing their fluorescence. Here, we show that AgNPs fed to cells to accumulate within lysosomes enhanced the fluorescence of lysosome-targeted Alexa488-conjugated dextran, BODIPY-cholesterol, and DQ-BSA. Moreover, AgNP increased the fluorescence of GFP fused to the cytosolic tail of LAMP1, showing that metal enhanced fluorescence can occur across the lysosomal membrane. The inclusion of AgNPs in lysosomes did not disturb lysosomal properties such as lysosomal pH, degradative capacity, autophagy and autophagic flux, and membrane integrity, though AgNP seemed to increase basal lysosome tubulation. Importantly, by using AgNP, we could track lysosome motility with reduced laser power without damaging and altering lysosome dynamics. Overall, AgNP-enhanced fluorescence may be a useful tool to study the dynamics of the endo-lysosomal pathway while minimizing phototoxicity. The American Society for Cell Biology 2023-08-17 /pmc/articles/PMC10551705/ /pubmed/37405751 http://dx.doi.org/10.1091/mbc.E22-06-0200 Text en © 2023 Soha et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 4.0 Unported Creative Commons License. |
spellingShingle | Special Issue on Cell Biology of the Lysosome Soha, Sumaiya A. Santhireswaran, Araniy Huq, Saaimatul Casimir-Powell, Jayde Jenkins, Nicala Hodgson, Gregory K. Sugiyama, Michael Antonescu, Costin N. Impellizzeri, Stefania Botelho, Roberto J. Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title | Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title_full | Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title_fullStr | Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title_full_unstemmed | Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title_short | Improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
title_sort | improved imaging and preservation of lysosome dynamics using silver nanoparticle-enhanced fluorescence |
topic | Special Issue on Cell Biology of the Lysosome |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10551705/ https://www.ncbi.nlm.nih.gov/pubmed/37405751 http://dx.doi.org/10.1091/mbc.E22-06-0200 |
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