A simplified method for immunostaining single endometrial and liver organoids

Organoids derived from tissues such as the endometrium and liver represent crucial models for investigating cellular and molecular processes in a three-dimensional framework. These systems enable detailed studies of organ development and disease pathology through various methods, including immunostaining, for precise visualization and localization of specific proteins. This simplified method outlines a systematic protocol for isolating, culturing, and immunostaining individual organoids using transwell inserts. This streamlined method optimizes procedures for fixation, permeabilization, and single or double antibody staining, ensuring robust and reproducible outcomes. Our method allows selecting a single organoid in a temporal manner without disturbing the 3D structure of Matrigel domes. By standardizing these critical steps, researchers can efficiently utilize organoid cultures to advance our understanding of complex biological mechanisms and explore therapeutic interventions targeting diseases affecting the endometrium and liver.