Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM)

Irina A. Okkelman; Hang Zhou; Sergey M. Dmitriev; Angela C. Debruyne; Austin E. Y. T. Lefebvre; Marcelo Leomil Zoccoler; Linglong Chen; Bert Devriendt; Ruslan I. Dmitriev

doi:10.1038/s41377-025-01949-0

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Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM)

Original Articles

- Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM)
- Light: Science & Applications Vol. 14, Issue 10, Pages: 2915-2935(2025)
- Affiliations：
  
  1.Tissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, The Core, C. Heymanslaan 10, 9000 Ghent, Belgium
  2.Ghent Light Microscopy Core, Ghent University, 9000 Ghent, Belgium
  3.Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, Graz 8010, Austria
  4.Calico Life Sciences LLC, South San Francisco, CA 94080, USA
  5.BioImage Analysis Technology Development Group, DFG Cluster of Excellence “Physics of Life”, TU Dresden, 01307 Dresden, Germany
  6.Laboratory of Immunology, Department of Translational Physiology, Infectiology and Public Health, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium
- Author bio：
  
  Bert Devriendt (b.devriendt@ugent.be)
  Ruslan I. Dmitriev (Ruslan.dmitriev@ugent.be)
- Funds：
- DOI：10.1038/s41377-025-01949-0
  CLC：
- Received：12 February 2025，
  
  Revised：2025-06-25，
  
  Accepted：01 July 2025，
  
  Published Online：12 August 2025，
  
  Published：31 October 2025
- Accepted：
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Okkelman, I. A. et al. Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM). Light: Science & Applications, 14, 2915-2935 (2025).
DOI：

Okkelman, I. A. et al. Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM). Light: Science & Applications, 14, 2915-2935 (2025). DOI： 10.1038/s41377-025-01949-0.

摘要

Abstract

Increased micro- and nanoplastic (MNP) pollution poses significant health risks

yet the mechanisms of their accumulation and effects on absorptive tissues remain poorly understood. Addressing this knowledge gap requires tractable models coupled to dynamic live cell imaging methods

enabling multi-parameter single cell analysis. We report a new method combining adult stem cell-derived small intestinal organoid cultures with live fluorescence lifetime imaging microscopy (FLIM) to study MNP interactions with gut epithelium. To facilitate this

we optimized live imaging of porcine and mouse small intestinal organoids with an ‘apical-out’ topology. Subsequently

we produced a set of pristine MNPs based on PMMA and PS (<200 nm

doped with deep-red fluorescent dye) and evaluated their interaction with organoids displaying controlled epithelial polarity. We found that nanoparticles interacted differently with apical and basal membranes of the organoids and showed a species-specific pattern of cellular uptake. Using a phasor analysis approach

we demonstrate improved sensitivity of FLIM over conventional intensity-based microscopy. The resulting ‘fluorescence lifetime barcoding’ enabled distinguishing of different types of MNP and their interaction sites within organoids. Finally

we studied short (1 day)- and long (3 day)-term exposure effects of PMMA and PS-based MNPs on mitochondrial function

total cell energy budget and epithelial inflammation. We found that even pristine MNPs could disrupt chemokine production and mitochondrial membrane potential in intestinal epithelial cells. The presented FLIM approach will advance the study of MNP toxicity

their biological impacts on gastrointestinal tissue and enable the tracing of other fluorescent nanoparticles in live organoid and 3D ex vivo systems.

关键词

Keywords

references

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