Detection and Quantification of Nile Red Stained Microplastic Particulates in Breastmilk using Fiji

Microplastics are small, water-insoluble plastic particles that measure between 1 micrometer and 1 millimeter in size, and have been found in various environmental matrices. Their small size allows them to infiltrate cells and even nuclei, posing potential risks to human health through mechanisms such as oxidative damage, DNA alteration, and disruption of biological processes. Despite their widespread presence, there is a notable gap in research on the presence of microplastics in human breast milk, with only a few studies having investigated this area of study. Recently, advanced identification is being used to detect and analyze microplastics in various environments, over traditional techniques such as visual sorting, sieving, and density separation. In this study, a modified staining technique was developed for detecting microplastics in breast milk, utilizing a combination of fluorescent staining and Fenton's reagent that was specifically selected to be digest the fats, sugars, and proteins that are found in breast milk. Quantitative analysis of microplastics was achieved via the development of a custom image processing protocol using Fiji software, which utilizes programming scripts to automate data analysis. Our findings indicate that microplastics may be present in breastmilk, however, limitations in our current methodology, particularly the inaccessibility of advanced equipment, hinder our ability to precisely identify microplastic particulates based on chemical composition Future research could benefit from utilizing techniques including Raman spectroscopy, which may offer enhanced precision in detecting and quantifying microplastics in human breast milk. This study highlights the need for further in-depth investigation to better understand the implications of microplastic presence in breastmilk.