Analytical and Bioanalytical Chemistry

, Volume 411, Issue 16, pp 3497–3506 | Cite as

High-resolution laser ablation inductively coupled plasma mass spectrometry used to study transport of metallic nanoparticles through collagen-rich microstructures in fibroblast multicellular spheroids

  • Akihiro ArakawaEmail author
  • Norbert Jakubowski
  • Sabine Flemig
  • Gunda Koellensperger
  • Mate Rusz
  • Daigo Iwahata
  • Heike Traub
  • Takafumi Hirata
Research Paper


We have efficiently produced collagen-rich microstructures in fibroblast multicellular spheroids (MCSs) as a three-dimensional in vitro tissue analog to investigate silver (Ag) nanoparticle (NP) penetration. The MCS production was examined by changing the seeding cell number (500 to 40,000 cells) and the growth period (1 to 10 days). MCSs were incubated with Ag NP suspensions with a concentration of 5 μg mL−1 for 24 h. For this study, laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used to visualize Ag NP localization quantitatively. Thin sections of MCSs were analyzed by LA-ICP-MS with a laser spot size of 8 μm to image distributions of 109Ag, 31P, 63Cu, 66Zn, and 79Br. A calibration using a NP suspension was applied to convert the measured Ag intensity into the number of NPs present. The determined numbers of NPs ranged from 30 to 7200 particles in an outer rim of MCS. The particle distribution was clearly correlated with the presence of 31P and 66Zn and was localized in the outer rim of proliferating cells with a width that was equal to about twice the diameter of single cells. Moreover, abundant collagens were found in the outer rim of MCSs. For only the highest seeding cell number, NPs were completely captured at the outer rim, in a natural barrier reducing particle transport, whereas Eosin (79Br) used as a probe of small molecules penetrated into the core of MCSs already after 1 min of exposure.

Graphical abstract

Fibroblast MCS could build up the barrier only for nanoparticles


Laser ablation inductively coupled plasma mass spectrometry Silver nanoparticles Fibroblast cells Multicellular spheroids 



We thank Konrad Löhr (Bundesanstalt für Materialforschung und -prüfung) for the support and training for the non-contact piezo-driven array spotter and Akvile Häckel (Charité Universitätsmedizin Berlin) for providing access to and support with using the cryomicrotome.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2019_1827_MOESM1_ESM.pdf (1.2 mb)
ESM 1 (PDF 1191 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Bundesanstalt für Materialforschung und -prüfung (BAM)BerlinGermany
  2. 2.Research Institute for Bioscience Products and Fine ChemicalsAjinomoto Co., Inc.Kawasaki-shiJapan
  3. 3.Spetec GmbHErdingGermany
  4. 4.Institute of Analytical ChemistryUniversity of ViennaViennaAustria
  5. 5.Cell Culture Facility, Institute of Inorganic ChemistryUniversity of ViennaViennaAustria
  6. 6.Geochemical Research CenterThe University of TokyoTokyoJapan

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