Underappreciated microplastic galaxy biases the filter-based quantification

Long-term environmental loading of microplastics (MPs) causes alarming exposure risks for a variety of species worldwide, considered a planetary threat to the well-being of ecosystems. Robust quantitative estimates of MP extents and featured diversity are the basis for comprehending their environmental implications precisely, and of these methods, membrane-based characterizations predominate with respect to MP inspections. However, though crucial to filter-based MP quantification, aggregation statuses of retained MPs on these substrates remain poorly understood, leaving us a "blind box" that exaggerates uncertainty in quantitive strategies of preselected areas without knowing overview loading structure. To clarify this uncertainty and estimate their impacts on MP counting, using MP imaging data assembled from peer-reviewed studies through a systematic review, here we analyze the particle-specific profiles of MPs retained on various substrates according to their centre of mass with a fast-random forests algorithm. We visualize the formation of distinct galaxy-like MP aggregation-similar to the solar system and Milky Way System comprised of countless stars-across the pristine and environmental samples by leveraging two spatial parameters developed in this study. This unique pattern greatly challenges the homogeneously or randomly distributed MP presumption adopted extensively for simplified membrane-based quantification purposes and selective ROI (region of interest) estimates for smaller-sized plastics down to the nano-range, as well as the compatibility theory using pristine MPs as the standard to quantify the presence of environmental MPs. Furthermore, our evaluation with exemplified numeration cases confirms these location-specific and area-dependent biases in many imaging analyses of a selective filter area, ascribed to the minimum possibility of reaching an ideal turnover point for the selective quantitive strategies. Consequently, disproportionate MP schemes on loading substrates yield great uncertainty in their quantification processing, highlighting the prompt need to include pattern-resolved calibration prior to quantification. Our findings substantially advance our understanding of the structure, behavior, and formation of these MP aggregating statuses on filtering substrates, addressing a fundamental question puzzling scientists as to why reproducible MP quantification is barely achievable even for subsamples. This study inspires the following studies to reconsider the impacts of aggregating patterns on the effective counting protocols and target-specific removal of retained MP aggregates through membrane separation techniques.