DeleteROI for Cleaning CiliaQ Output of Non-ciliary Contamination

MicroPubl Biol. 2025 Aug 14:2025:10.17912/micropub.biology.001670. doi: 10.17912/micropub.biology.001670. eCollection 2025.

Abstract

The ImageJ plugin CiliaQ developed by Hansen and colleagues (Hansen et al., 2021) provides for sophisticated analysis of ciliary parameters in three-dimensional space. However, midbodies and other non-ciliary structures can contaminate the output and require significant effort to remove. Furthermore, the manual removal of contamination risks subjective bias as the data is not blinded to the investigator. To address these problems, we developed an ImageJ plugin that presents images of the cilia region-of-interests (ROIs) identified by CiliaQ in a clickable grid that allows for marking and automated removal of non-ciliary contaminants. To reduce subjective bias, our plugin works on a dataset of multiple images and presents the cilia ROIs randomly. If the dataset contains both control and experimental conditions, the cilia are randomly interspersed with no visible information about their experimental group, thus reducing subjective bias. After removal of contamination, the cleaned data is output maintaining the CiliaQ file formats initially used.