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

Galluzzi，Massimiliano1; Zhang，Bokai2,3,4; Zhang，Han2,5; Wang，Lingzhi6; Lin，Yuan7,8; Yu，Xue Feng1; Chu，Zhiqin9; Li，Jiangyu2,10

2021-05-06

10.3389/fmolb.2021.669361

Frontiers in Molecular Biosciences   影响因子和分区

2296-889X

Fluorescent 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.

atomic force microscopy biomembrane correlative fluorescence microscopy fluorophore hybrid phospholipids

SCI

英语

其他

WOS:000652172100001

2-s2.0-85106159889

Scopus

1.Materials Interfaces Center,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen,China
2.Shenzhen Key Laboratory of Nanobiomechanics,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen,China
3.DGene (Dongjin Big Health (Shenzhen)) Co.,Ltd,Shenzhen,China
4.BenHealth Biopharmaceutical (Shenzhen) Co.,Ltd,Shenzhen,China
5.State Key Laboratory of Traction Power,Southwest Jiaotong Univerisity,Chengdu,China
6.Department of Electrical and Electronic Engineering,The University of Hong Kong,Hong Kong
7.Department of Mechanical Engineering,The University of Hong Kong,Hong Kong
8.Advanced Biomedical Instrumentation Centre,Shatin,Hong Kong
9.Department of Electrical and Electronic Engineering,Joint Appointment with School of Biomedical Sciences,The University of Hong Kong,Hong Kong
10.Department of Materials Science and Engineering,Southern University of Science and Technology,Shenzhen,China

Galluzzi，Massimiliano,Zhang，Bokai,Zhang，Han,et al. Unveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis[J]. Frontiers in Molecular Biosciences,2021,8.

Galluzzi，Massimiliano.,Zhang，Bokai.,Zhang，Han.,Wang，Lingzhi.,Lin，Yuan.,...&Li，Jiangyu.(2021).Unveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis.Frontiers in Molecular Biosciences,8.

Galluzzi，Massimiliano,et al."Unveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis".Frontiers in Molecular Biosciences 8(2021).