Cell-Nanoparticle Interactions at (Sub)-Nanometer Resolution Analyzed by Electron Microscopy and Correlative Coherent Anti-Stokes Raman Scattering

Jukka Kalle Samuel Saarinen, Friederike Gütter, Mervi M Lindman, Mikael Agopov, Sara J. Fraser-Miller, Regina Scherließ, Eija Jokitalo, Helder Almeida Santos, Leena Peltonen, Antti Isomäki, Clare J. Strachan

Research output: Contribution to journalArticleScientificpeer-review

Abstract

A wide variety of nanoparticles are playing an increasingly important role in drug delivery. Label-free imaging techniques are especially desirable to follow the cellular uptake and intracellular fate of nanoparticles. The combined correlative use of different techniques, each with unique advantages, facilitates more detailed investigation about such interactions. The synergistic use of correlative coherent anti-Stokes Raman scattering and electron microscopy (C-CARS-EM) imaging offers label-free, chemically-specific, and (sub)-nanometer spatial resolution for studying nanoparticle uptake into cells as demonstrated in the current study. Coherent anti-Stokes Raman scattering (CARS) microscopy offers chemically-specific (sub)micron spatial resolution imaging without fluorescent labels while transmission electron microscopy (TEM) offers (sub)-nanometer scale spatial resolution and thus visualization of precise nanoparticle localization at the sub-cellular level. This proof-of-concept imaging platform with unlabeled drug nanocrystals and macrophage cells revealed good colocalization between the CARS signal and electron dense nanocrystals in TEM images. The correlative TEM images revealed subcellular localization of nanocrystals inside membrane bound vesicles, showing multivesicular body (MVB)-like morphology typical for late endosomes (LEs), endolysosomes, and phagolysosomes. C-CARS-EM imaging has much potential to study the interactions between a wide range of nanoparticles and cells with high precision and confidence.

Original languageEnglish
Article number1800413
JournalBiotechnology Journal
Volume14
Issue number4
Number of pages10
ISSN1860-6768
DOIs
Publication statusPublished - Apr 2019
MoE publication typeA1 Journal article-refereed

Fields of Science

  • FLUORESCENCE
  • IN-VITRO
  • LASER
  • LOCALIZATION
  • SPECTRA
  • SPECTROSCOPY
  • TOOL
  • cell imaging
  • cellular nanoparticle uptake
  • coherent anti-Stokes Raman scattering (CARS) microscopy
  • drug nanocrystals
  • non-linear imaging
  • 1182 Biochemistry, cell and molecular biology
  • 116 Chemical sciences
  • 317 Pharmacy

Equipment

Biomedicum Imaging Unit

Elina Ikonen (Manager)

Medicum

Facility/equipment: Core Facility

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