Visualizing the internalization and biological impact of nanoplastics in live intestinal organoids by Fluorescence Lifetime Imaging Microscopy (FLIM)
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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.
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Sasa, G. B. K., Borlay, A. J., & Tan, C. S. (2026) · Journal of Electroanalytical Chemistry · doi:10.1016/j.jelechem.2026.120310
Xiftou, K., Jiang, J., Caiment, F., de Kok, T. M. C. M., Vinken, M., Sanz-Serrano, J., & Tabernilla, A. (2026) · Archives of Toxicology · doi:10.1007/s00204-026-04473-1
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Fate, toxicology and Organ-on-a-chip methodology of nanoplastics in human body
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Pathogenesis of Micro/Nanoplastics in Mammalian Systems: Gut to Systemic Multi-Organ Dysfunction
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Nanoplastics reshape local intracellular RNP environments and P-body dynamicsPreprint
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Fluorescent Labeling Strategies for Tracking Micro- and Nanoplastics in Biological Systems
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Monitoring Calcium Dynamics in Pig Intestinal Organoids with FLIM and FRET
Cervellera, M., Zhou, H., van den Broek, B., Jalink, K., & Dmitriev, R. I. (2026) · Methods in molecular biology · doi:10.1007/978-1-0716-5412-5_17
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Quantifying Cellular Nanoplastic Accumulation Using Raman Microscopy
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- Authors
- Okkelman, I. A., Zhou, H., Borisov, S. M., Debruyne, A. C., Lefebvre, A. E. Y. T., Leomil Zoccoler, M., Chen, L., Devriendt, B., & Dmitriev, R. I.
- Affiliations
- Ghent University (BE)
- Graz University of Technology (AT)
- Calico Life Sciences (US)
- Corresponding author
- Devriendt, Bert
- Dmitriev, Ruslan I.
- ORCID
- Venue
- Light: Science & Applications
- Published
- 2025-08-12
- Volume
- 14
- Issue
- 1
- Pages
- 272-272
- ISSN
- 2047-7538 (ISSN-L)
- PMC
- PMC12343940
- OpenAlex
- W4413111363
- Cited by
- 28
- References
- 78
- Citations by year
- 2026: 20 · 2025: 8
- FWCI
- 6.60
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Light Science & Applications (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion, cc-by-nc-nd)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- EC | Horizon 2020 Framework Programme (101073507)
- Topics
- Physical Sciences › Environmental Science › Pollution › Microplastics and Plastic Pollution
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Orbital Angular Momentum in Optics
- Keywords
- nanoplastics, microplastics, intestinal organoids, small intestine, fluorescence lifetime imaging, phasor analysis, nanoparticle uptake, epithelial polarity, mitochondrial function, mitochondrial membrane potential, chemokine production, epithelial inflammation, energy metabolism, PMMA nanoparticles, polystyrene nanoparticles, nanoplastic toxicity, live-cell imaging
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