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Article: Unveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis

TitleUnveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis
Authors
Keywordsatomic force microscopy
correlative fluorescence microscopy
biomembrane
fluorophore
hybrid phospholipids
Issue Date2021
PublisherFrontiers Research Foundation. The Journal's web site is located at https://www.frontiersin.org/journals/molecular-biosciences
Citation
Frontiers in Molecular Biosciences, 2021, v. 8, p. article no. 669361 How to Cite?
AbstractFluorescent imaging combined with atomic force microscopy (AFM), namely AFM-fluorescence correlative microscopy, is a popular technology in life science. However, the influence of involved fluorophores on obtained mechanical information is normally underestimated, and such subtle changes are still challenging to detect. Herein, we combined AFM with laser light excitation to perform a mechanical quantitative analysis of a model membrane system labeled with a commonly used fluorophore. Mechanical quantification was additionally validated by finite element simulations. Upon staining, we noticed fluorophores forming a diffuse weakly organized overlayer on phospholipid supported membrane, easily detected by AFM mechanics. The laser was found to cause a degradation of mechanical stability of the membrane synergically with presence of fluorophore. In particular, a 30 min laser irradiation, with intensity similar to that in typical confocal scanning microscopy experiment, was found to result in a ∼40% decrease in the breakthrough force of the stained phospholipid bilayer along with a ∼30% reduction in its apparent elastic modulus. The findings highlight the significance of analytical power provided by AFM, which will allow us to “see” the “unseen” in correlative microscopy, as well as the necessity to consider photothermal effects when using fluorescent dyes to investigate, for example, the deformability and permeability of phospholipid membranes.
Persistent Identifierhttp://hdl.handle.net/10722/305352
ISSN
2023 Impact Factor: 3.9
2023 SCImago Journal Rankings: 1.232
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorGalluzzi, M-
dc.contributor.authorZhang, B-
dc.contributor.authorZhang, H-
dc.contributor.authorWang, L-
dc.contributor.authorLin, Y-
dc.contributor.authorYu, XF-
dc.contributor.authorChu, Z-
dc.contributor.authorLi, J-
dc.date.accessioned2021-10-20T10:08:12Z-
dc.date.available2021-10-20T10:08:12Z-
dc.date.issued2021-
dc.identifier.citationFrontiers in Molecular Biosciences, 2021, v. 8, p. article no. 669361-
dc.identifier.issn2296-889X-
dc.identifier.urihttp://hdl.handle.net/10722/305352-
dc.description.abstractFluorescent imaging combined with atomic force microscopy (AFM), namely AFM-fluorescence correlative microscopy, is a popular technology in life science. However, the influence of involved fluorophores on obtained mechanical information is normally underestimated, and such subtle changes are still challenging to detect. Herein, we combined AFM with laser light excitation to perform a mechanical quantitative analysis of a model membrane system labeled with a commonly used fluorophore. Mechanical quantification was additionally validated by finite element simulations. Upon staining, we noticed fluorophores forming a diffuse weakly organized overlayer on phospholipid supported membrane, easily detected by AFM mechanics. The laser was found to cause a degradation of mechanical stability of the membrane synergically with presence of fluorophore. In particular, a 30 min laser irradiation, with intensity similar to that in typical confocal scanning microscopy experiment, was found to result in a ∼40% decrease in the breakthrough force of the stained phospholipid bilayer along with a ∼30% reduction in its apparent elastic modulus. The findings highlight the significance of analytical power provided by AFM, which will allow us to “see” the “unseen” in correlative microscopy, as well as the necessity to consider photothermal effects when using fluorescent dyes to investigate, for example, the deformability and permeability of phospholipid membranes.-
dc.languageeng-
dc.publisherFrontiers Research Foundation. The Journal's web site is located at https://www.frontiersin.org/journals/molecular-biosciences-
dc.relation.ispartofFrontiers in Molecular Biosciences-
dc.rightsThis Document is Protected by copyright and was first published by Frontiers. All rights reserved. It is reproduced with permission.-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.subjectatomic force microscopy-
dc.subjectcorrelative fluorescence microscopy-
dc.subjectbiomembrane-
dc.subjectfluorophore-
dc.subjecthybrid phospholipids-
dc.titleUnveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis-
dc.typeArticle-
dc.identifier.emailChu, Z: zqchu@eee.hku.hk-
dc.identifier.authorityChu, Z=rp02472-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.3389/fmolb.2021.669361-
dc.identifier.pmid34026842-
dc.identifier.pmcidPMC8136518-
dc.identifier.scopuseid_2-s2.0-85106159889-
dc.identifier.hkuros328151-
dc.identifier.volume8-
dc.identifier.spagearticle no. 669361-
dc.identifier.epagearticle no. 669361-
dc.identifier.isiWOS:000652172100001-
dc.publisher.placeSwitzerland-

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