Recent advances in the standardization of fluorescence microscopy for quantitative image analysis.

Fluorescence microscopy is a broadly used technique within a variety of different fields, including life sciences and medical research, and its quantitative aspects are becoming emphasized. The challenge now lies in improving the accuracy and precision of the measurements obtained from fluorescence microscopy. Improving these will facilitate the comparison of results between different instruments/institutions and therefore ensure the reproducibility of results. In this review, recent standardization procedures, including the benchmarking of the instrument performance and the standardization of the image itself, as well as the reference materials for calibration are summarized, and an overview of the advances in fluorescence microscopy standardization and the current limitations are presented. A procedure for the comparison of the image data obtained using different instruments, by different analysts and/or at different times, should be developed to improve the standardization of this data. The standardization of image data would lead to the development of new applications of fluorescence microscopy not only in academic research but also in regulatory science. © International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature 2021.