Wastewater effluent is expected to be a pathway for microplastics to enter the aquatic environment, with microbeads from cosmetic products and polymer fibres from clothes likely to enter wastewater treatment plants (WWTP). To date, few studies have quantified microplastics in wastewater. Moreover, the lack of a standardized and applicable method to identify microplastics in complex samples, such as wastewater, has limited the accurate assessment of microplastics and may lead to an incorrect estimation. This study aimed to develop a validated method to sample and process microplastics from wastewater effluent and to apply the developed method to quantify and characterise wastewater-based microplastics in effluent from three WWTPs that use primary, secondary and tertiary treatment processes. We applied a high-volume sampling device that fractionated microplastics in situ and an efficient sample processing procedure to improve the sampling of microplastics in wastewater and to minimize the false detection of non-plastic particles. The sampling device captured between 92% and 99% of polystyrene microplastics using 25 μm–500 μm mesh screens in laboratory tests. Microplastic type, size and suspected origin in all studied WWTPs, along with the removal efficiency during the secondary and tertiary treatment stages, was investigated. Suspected microplastics were characterised using Fourier Transform Infrared spectroscopy, with between 22 and 90% of the suspected microplastics found to be non-plastic particles. An average of 0.28, 0.48 and 1.54 microplastics per litre of final effluent was found in tertiary, secondary and primary treated effluent, respectively. This study suggests that although low concentrations of microplastics are detected in wastewater effluent, WWTPs still have the potential to act as a pathway to release microplastics given the large volumes of effluent discharged to the aquatic environment. This study focused on a single sampling campaign, with long-term monitoring recommended to further characterise microplastics in wastewater.