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Automated quantification of fluorescence signals in bivalve hemocytes (Mytilus edulis) as a proxy for micro-and-nanoplastic toxicity assessment

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Abstract
Plastic particles are prevalent in the environment, and the risks they pose especially to aquatic organisms are of growing concern. A range of responses attributed to micro-and-nanoplastic particle (MPs/NPs) exposure has been documented across various levels of biological organization, extending from cellular to population-level endpoints. However, systematic analyses of effect endpoints at the cellular level, which are considered a sensitive response variable to plastic particle exposure, are still lacking. It has been established that plastic particles can be internally distributed in organisms through various ways, such as translocation and penetration of biological barriers. In particular, particles with dimensions in the order of nanometers (i.e. nanoplastics, NPs, <1000 nm) are able to enter cells via endocytotic pathways (i.e. phagocytosis). These cellular processes have been highlighted as integral factors that influence key physiological functions including cellular immunity. Hemocytes are responsible for cell-mediated immunity in bivalve mollusks such as the mussel Mytilus edulis, which is highly susceptible to NP exposure due to its sedentary and filter-feeding lifestyle. The hemocytes circulate within the hemolymph and can cross all epithelial boundaries, acting as phagocytes against foreign particles, including plastic particles. Assessing the different physicochemical properties of MPs/NPs, such as polymer type, size, shape, and surface charge, is crucial as these parameters may directly affect the cellular uptake mechanisms as well as cytotoxicity. Hence, this study aims to validate and apply a microscopy-based method for determining whether MPs/NPs exposure induces apoptosis/necrosis (cellular death) in the hemocyte of mussels M. edulis as a proxy for micro-and-nanoplastic risk assessment. An in-vitro approach will be applied to characterize and interpret the response profiles under environmentally relevant exposure conditions and in assessing differential effects linked to characteristics of the plastic particles (type, size, shape). The apoptosis and necrosis signal quantification will be carried out through dual staining approach using an apoptosis detection kit (Biovision) and by the complementary application of fluorescence microscopy and ImageJ, an open-source image processing program. The results will provide insights into the underlying mechanisms associated with different physicochemical characteristics of MPs/NPs at a cellular level which is crucial for fate and toxicity assessment in aquatic organisms.
Keywords
hemocytes, micro-and-nanoplastics, immunotoxicity, microscopy, ImageJ

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MLA
Hara, Jenevieve, et al. “Automated Quantification of Fluorescence Signals in Bivalve Hemocytes (Mytilus Edulis) as a Proxy for Micro-and-Nanoplastic  Toxicity Assessment.” Interfaces Against Pollution 2022, Abstracts, 2022.
APA
Hara, J., Schoenaers, S., Vercauteren, M., Janssen, C., Blust, R., Asselman, J., & Town, R. (2022). Automated quantification of fluorescence signals in bivalve hemocytes (Mytilus edulis) as a proxy for micro-and-nanoplastic  toxicity assessment. Interfaces Against Pollution 2022, Abstracts. Presented at the Interfaces Against Pollution: Chemical and Biological Perspectives (IAP 2022), Antwerp, Belgium.
Chicago author-date
Hara, Jenevieve, Sébastjen Schoenaers, Maaike Vercauteren, Colin Janssen, Ronny Blust, Jana Asselman, and Raewyn Town. 2022. “Automated Quantification of Fluorescence Signals in Bivalve Hemocytes (Mytilus Edulis) as a Proxy for Micro-and-Nanoplastic  Toxicity Assessment.” In Interfaces Against Pollution 2022, Abstracts.
Chicago author-date (all authors)
Hara, Jenevieve, Sébastjen Schoenaers, Maaike Vercauteren, Colin Janssen, Ronny Blust, Jana Asselman, and Raewyn Town. 2022. “Automated Quantification of Fluorescence Signals in Bivalve Hemocytes (Mytilus Edulis) as a Proxy for Micro-and-Nanoplastic  Toxicity Assessment.” In Interfaces Against Pollution 2022, Abstracts.
Vancouver
1.
Hara J, Schoenaers S, Vercauteren M, Janssen C, Blust R, Asselman J, et al. Automated quantification of fluorescence signals in bivalve hemocytes (Mytilus edulis) as a proxy for micro-and-nanoplastic  toxicity assessment. In: Interfaces Against Pollution 2022, Abstracts. 2022.
IEEE
[1]
J. Hara et al., “Automated quantification of fluorescence signals in bivalve hemocytes (Mytilus edulis) as a proxy for micro-and-nanoplastic  toxicity assessment,” in Interfaces Against Pollution 2022, Abstracts, Antwerp, Belgium, 2022.
@inproceedings{8767570,
  abstract     = {{Plastic particles are prevalent in the environment, and the risks they pose especially to aquatic organisms are of growing concern. A range of responses attributed to micro-and-nanoplastic particle (MPs/NPs) exposure has been documented across various levels of biological organization, extending from cellular to population-level endpoints. However, systematic analyses of effect endpoints at the cellular level, which are considered a sensitive response variable to plastic particle exposure, are still lacking. It has been established that plastic particles can be internally distributed in organisms through various ways, such as translocation and penetration of biological barriers. In particular, particles with dimensions in the order of nanometers (i.e. nanoplastics, NPs, <1000 nm) are able to enter cells via endocytotic pathways (i.e. phagocytosis). These cellular processes have been highlighted as integral factors that influence key physiological functions including cellular immunity. Hemocytes are responsible for cell-mediated immunity in bivalve mollusks such as the mussel Mytilus edulis, which is highly susceptible to NP exposure due to its sedentary and filter-feeding lifestyle. The hemocytes circulate within the hemolymph and can cross all epithelial boundaries, acting as phagocytes against foreign particles, including plastic particles. Assessing the different physicochemical properties of MPs/NPs, such as polymer type, size, shape, and surface charge, is crucial as these parameters may directly affect the cellular uptake mechanisms as well as cytotoxicity. Hence, this study aims to validate and apply a microscopy-based method for determining whether MPs/NPs exposure induces apoptosis/necrosis (cellular death) in the hemocyte of mussels M. edulis as a proxy for micro-and-nanoplastic risk assessment. An in-vitro approach will be applied to characterize and interpret the response profiles under environmentally relevant exposure conditions and in assessing differential effects linked to characteristics of the plastic particles (type, size, shape). The apoptosis and necrosis signal quantification will be carried out through dual staining approach using an apoptosis detection kit (Biovision) and by the complementary application of fluorescence microscopy and ImageJ, an open-source image processing program. The results will provide insights into the underlying mechanisms associated with different physicochemical characteristics of MPs/NPs at a cellular level which is crucial for fate and toxicity assessment in aquatic organisms.}},
  author       = {{Hara, Jenevieve and Schoenaers, Sébastjen and Vercauteren, Maaike and Janssen, Colin and Blust, Ronny and Asselman, Jana and Town, Raewyn}},
  booktitle    = {{Interfaces Against Pollution 2022, Abstracts}},
  keywords     = {{hemocytes,micro-and-nanoplastics,immunotoxicity,microscopy,ImageJ}},
  language     = {{eng}},
  location     = {{Antwerp, Belgium}},
  title        = {{Automated quantification of fluorescence signals in bivalve hemocytes (Mytilus edulis) as a proxy for micro-and-nanoplastic  toxicity assessment}},
  year         = {{2022}},
}