Pictures adapted from Biorender.com. PROJECT OUTLINE STABILITY AND DEGRADATION TESTS RECOVERY AND QUANTIFICATION OF SPIKED MNPs Fluorescent polystyrene (FPS) MNPs were recovered after digestion and filtration with Confocal Fluorescence Microscopy (CFM) and Scanning Electron Microscopy (SEM). No changes in morphology were observed. PLACENTAL TISSUE DIGESTION PROTOCOLS Spiked placental tissue was homogenized and enzymatically digested, followed by filtration over 700 nm glass fiber filters. Cleaner filter better digestion. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under AURORA grant agreement No 964827. LEARN MORE AND FOLLOW OUR PROGRESS QUESTIONS? COMMENTS? AuroraResearch.eu/newsletter Learn more about AURORA, the team mem- Do you have questions about AURORA? bers, and our latest research progress. Visit Want to engage with us or inquire about our website, follow our social media channels, an opportunity to collaborate? Then send and subscribe to our newsletter: us an email or get in touch directly with a responsible team member! AuroraResearch.eu info@AuroraResearch.eu @AuroraProjectEU AuroraResearch.eu/about/team linkedin.com/company/AuroraResearch NEWS Sample Pre-processing Protocols for the (Spectro-)Microscopic Analysis of Micro- & Nanoplastics (MNPs) in Maternal and Fetal Tissues Laura M. Zoutendijk1, Laurens D. B. Mandemaker1, Zenzi Matla1, Lisa Zimmermann2, Lonneke van Dalen1, Bert M. Weckhuysen1, Florian Meirer1 1: Inorganic Chemistry and Catalysis, Debye Institute for Nanomaterials Science and Institute for Sustainable and Circular Chemistry, Utrecht University, Universiteitsweg 99, 3584 CG Utrecht, The Netherlands 2: Food Packaging Forum, Staffelstrasse 10, CH-8045 Zurich, Switzerland Morphology Chemical composition Surface chemistry State of degradation Quantity Cultured spiked placenta cells • Optimize digestion • Ensure MNP stability Spiked placenta tissue • Optimize digestion and filtration • Find back MNPs • Determine recovery rate Non-spiked (environmental) placenta tissue • Detect MNPs • Determine MNP location • Analyze MNP degradation within body 200 nm FPS in water, 95 °C, 30 min. No agglomeration 200 nm FPS in 30% H2O2 No agglomeration 200 nm FPS in water, 95 °C, 2 hours Agglomeration occurs 200 nm FPS in pancreatin in TRIS-HCl No agglomeration Solution of badly digested tissue Homogenized placenta in buffer CFM image of 1 µm FPS (a) and SEM image of 200 nm FPS (b) recovered after digestion and filtration. CFM image of 0.5 μL droplet of 500 nm FPS in water (c) with color adjustments. Non-adjusted image is in right bottom corner. CONCLUSIONS AND OUTLOOK Fluo-PS stable in used digestive agents. Digestion protocol made for placental tissue. Fluo-PS in spiked digested tissue analyzed with CFM. • Calculate recovery rate of Fluo-PS in spiked digested tissue. • Adapt protocol to blood, amniotic fluid, urine, etc. • Towards analysis of non-spiked samples Placenta as received Solution of well digested tissue Glass fiber filter, well digested tissue Glass fiber filter, badly digested tissue Laura Zoutendijk l.m.zoutendijk@uu.nl 500 nm • To quantify the total number of recovered particles, the number of particles in a whole droplet can be counted. • Counting grids are used to stitch multiple microscopy images together. • The number of counted particles can then be compared to the number of particles theoretically present in the droplet: Recovery rate = Number of particles found back during analysis Theoretical number of spiked particles * 100% c b a