Correlating Super-Resolution Microscopy and Transmission Electron Microscopy Reveals Multiparametric Heterogeneity in Nanoparticles

Teodora Andrian, Pietro Delcanale, Silvia Pujals (Corresponding author), Lorenzo Albertazzi (Corresponding author)

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The functionalization of nanoparticles with functional moieties is a key strategy to achieve cell targeting in nanomedicine. The interplay between size and ligand number is crucial for the formulation performance and needs to be properly characterized to understand nanoparticle structure-activity relations. However, there is a lack of methods able to measure both size and ligand number at the same time and at the single particle level. Here, we address this issue by introducing a correlative light and electron microscopy (CLEM) method combining super-resolution microscopy (SRM) and transmission electron microscopy (TEM) imaging. We apply our super-resCLEM method to characterize the relationship between size and ligand number and density in PLGA-PEG nanoparticles. We highlight how heterogeneity found in size can impact ligand distribution and how a significant part of the nanoparticle population goes completely undetected in the single-technique analysis. Super-resCLEM holds great promise for the multiparametric analysis of other parameters and nanomaterials.

Original languageEnglish
Pages (from-to)5360–5368
Number of pages9
JournalNano Letters
Issue number12
Publication statusPublished - 14 Jun 2021


This work was supported by a Marie Skłodowska-Curie Horizon 2020 (713673), “la Caixa” Foundation (ID 100010434, code LCF/BQ/DI18/11660039) fellowship. S.P. and L.A acknowledge the financial support by the Spanish Ministry of Science and Innovation (PID2019-109450RB-I00/AEI/10.13039/501100011033), European Research Council/Horizon 2020 (ERC-StG-757397), “la Caixa” Foundation (ID 100010434), and by the Generalitat de Catalunya (through the CERCA program and 2017 SGR 01536). L.A. thanks the support of NWO through the VIDI Grant 192.028. The authors thank Lidia Delgado Valderrama and Maria Yolanda Muela Castro from the Electron Cryomicroscopy Unit from CCiTUB.

FundersFunder number
European Union 's Horizon 2020 - Research and Innovation Framework ProgrammeERC-StG-757397
Marie Skłodowska‐Curie
Marie Skłodowska‐Curie
“la Caixa” Foundation100010434, LCF/BQ/DI18/11660039
Horizon 2020 Framework Programme713673
Generalitat de Catalunya2017 SGR 01536
Nederlandse Organisatie voor Wetenschappelijk Onderzoek192.028
Ministerio de Ciencia e InnovaciónPID2019-109450RB-I00/AEI/10.13039/501100011033
Fundación Bancaria Caixa d'Estalvis i Pensions de Barcelona


    • correlative light and electron microscopy (CLEM)
    • electron microscopy (EM)
    • heterogeneity
    • nanomedicine
    • nanoparticles
    • super-resolution microscopy (SRM)
    • Nanoparticles
    • Microscopy, Electron, Transmission
    • Ligands
    • Microscopy, Fluorescence


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