Machine Learning‐Based Estimation of Experimental Artifacts and Image Quality in Fluorescence Microscopy

Reliable characterization of image data is fundamental for imaging applications, FAIR data management, and an objective evaluation of image acquisition, processing, and analysis steps in an image‐based investigation of biological samples. Image quality assessment (IQA) often relies on human visual perception, which is not objective, or reference ground truth images, which are not often available. This study presents a method for a comprehensive IQA of microscopic images, which solves these issues by employing a set of reference‐free metrics that estimate the presence of experimental artifacts. The metrics are jointly validated on a semisynthetic dataset and are tested on experimental images. Finally, the metrics are employed in a machine learning model, demonstrating their effectiveness for automatic artifact classification through multimarker IQA. This work provides a reliable reference‐free method for IQA in optical microscopy, which can be integrated into the experimental workflow and tuned to address specific artifact detection tasks.