Use of fluorescent-labelled nanoplastics (NPs) to demonstrate NP absorption is inconclusive without adequate controls

Catarino, Ana I.; Frutos, Amelie; Henry, Theodore B.

doi:10.1016/j.scitotenv.2019.03.194

Cited by 136 publications

(89 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Efforts to quantify PS-NP within mussel tissues was unsuccessful, as we were unable to distinguish the red fluorescence signal of the particles from autofluorescence. However, we remain confident that uptake occurred given our observations of active feeding, and evidence from other research papers that PS-NP (either free-floating or aggregated with microalgae) can be filtered-out by mussels; in these studies, PS-NP uptake was ascertained by measuring reduction of nanoparticle concentrations in the surrounding water and/or visualising uptake of PS-NP within tissues via fluorescence imaging (Brandts et al, 2018a;Sendra et al, 2019;Wegner et al, 2012), although such imaging studies have been criticised as leaching of fluorophores may give false positives (Catarino et al, 2019). identified per mg of digestive gland tissue of individual mussels, following 24 h or 7-day exposure to 500 ng mL -1 microplastics.…”

Section: Microplastic and Nanoplastic Uptakementioning

confidence: 96%

Microplastics, microfibres and nanoplastics cause variable sub-lethal responses in mussels (Mytilus spp.)

Cole

Liddle

Consolandi

et al. 2020

Marine Pollution Bulletin

163

View full text Add to dashboard Cite

We compare the toxicity of microplastics, microfibres and nanoplastics on mussels. Mussels (Mytilus spp.) were exposed to 500 ng mL-1 of 20 µm polystyrene microplastics, 10x30 µm polyamide microfibres or 50 nm polystyrene nanoplastics for 24 h or 7 days. Biomarkers of immune response, oxidative stress response, lysosomal destabilisation and genotoxic damage were measured in haemolymph, digestive gland and gills. Microplastics and microfibres were observed in the digestive glands, with significantly higher plastic concentrations after 7-days exposure (ANOVA, P<0.05). Nanoplastics had a significant effect on hyalinocyte-granulocyte ratios (ANOVA, P<0.05), indicative of a heightened immune response. SOD activity was significantly increased followed 24 h exposure to plastics (two-way ANOVA, P<0.05), but returned to normal levels after 7-days exposure. No evidence of lysosomal destabilisation or genotoxic damage was observed from any form of plastic. The study highlights how particle size is a key factor in plastic particulate toxicity.

show abstract

Section: Microplastic and Nanoplastic Uptakementioning

confidence: 96%

Microplastics, microfibres and nanoplastics cause variable sub-lethal responses in mussels (Mytilus spp.)

Cole

Liddle

Consolandi

et al. 2020

Marine Pollution Bulletin

163

View full text Add to dashboard Cite

show abstract

“…Fluorescently-labelled nanoplastics could leach fluorophores, since they are not covalently linked to the PS-NPs. 19 Catarino et al 20 suggested careful testing of the fluorescent particle uptake in order to avoid wrong conclusions. To establish the true internal accumulation of fluorescent PS-NPs, the following experiment was set up: 1 ml of PS-NP suspension (1000, 100 and 50 mg l −1 ) was centrifuged at 50 000 × g for 60 min, then the supernatant (SN) was transferred to a new Eppendorf tube and the pellet (P) resuspended in 1 ml of PBS.…”

Section: Decay Of Ps-np Fluorescence In Vitro and In Cell Culturementioning

confidence: 99%

Polystyrene nanoplastics accumulate in ZFL cell lysosomes and in zebrafish larvae after acute exposure, inducing a synergistic immune response in vitro without affecting larval survival in vivo

Brandts

Garcia-Ordoñez

Tort

et al. 2020

Environ. Sci.: Nano

View full text Add to dashboard Cite

show abstract

“…In the present study, our passive sampling approach could be an effective screening method for the stability of dyes in commercial nano-and microplastics. Very recently, Catarino et al (2019) published a study on zebrafish supporting the conclusion that dye leaching from 500 and 1000 nm polystyrene nanobeads causes artifacts. They suggested a dialysis step to remove uncoupled dye before performing toxicity studies.…”

Section: Dye Leaching Experiments (Experiments 4)mentioning

confidence: 96%

When Fluorescence Is not a Particle: The Tissue Translocation of Microplastics in Daphnia magna Seems an Artifact

Schür

Rist

Baun

et al. 2019

Enviro Toxic and Chemistry

145

View full text Add to dashboard Cite

Previous research reported the translocation of nano‐ and microplastics from the gastrointestinal tract to tissues in Daphnia magna, most prominently of fluorescent polystyrene beads to lipid droplets. For particles >300 nm, such transfer is biologically implausible as the peritrophic membrane retains these in the daphnid gut. We used confocal laser scanning microscopy to study tissue transfer applying the setup from a previous study (neonates exposed to 20 and 1000 nm polystyrene beads at 2 µg L–1 for 4 and 24 h), the same setup with a fructose‐based clearing, and a setup with a 1000‐fold higher concentration (2 mg L–1). We used passive sampling to investigate whether the beads leach the fluorescent dye. Although the 1000 nm beads were visible in the gut at both exposure concentrations, the 20 nm beads were detectable at 2 mg L–1 only. At this concentration, we observed fluorescence in lipid droplets in daphnids exposed to both particle types. However, this did not colocalize with the 1000 nm beads, which remained visible in the gut. We further confirmed the leaching of the fluorescent dye using a passive sampler, a method that can also be applied in future studies. In summary, we cannot replicate the original study but demonstrate that the fluorescence in the lipid droplets of D. magna results from leaching of the dye. Thus, the use of fluorescence as a surrogate for particles can lead to artifacts in uptake and translocation studies. This highlights the need to confirm the stability of the fluorescence label or to localize particles using alternative methods. Environ Toxicol Chem 2019;38:1495–1503. © 2019 SETAC Open Practices The present study has earned Open Data/Materials badges for making publicly available the digitally shareable data necessary to reproduce the reported results. Learn more about the Open Practices badges from the Center for Open Science: https://osf.io/tvyxz/wiki.

show abstract

Use of fluorescent-labelled nanoplastics (NPs) to demonstrate NP absorption is inconclusive without adequate controls

Cited by 136 publications

References 27 publications

Microplastics, microfibres and nanoplastics cause variable sub-lethal responses in mussels (Mytilus spp.)

Microplastics, microfibres and nanoplastics cause variable sub-lethal responses in mussels (Mytilus spp.)

Polystyrene nanoplastics accumulate in ZFL cell lysosomes and in zebrafish larvae after acute exposure, inducing a synergistic immune response in vitro without affecting larval survival in vivo

When Fluorescence Is not a Particle: The Tissue Translocation of Microplastics in Daphnia magna Seems an Artifact

Contact Info

Product

Resources

About