Live-cell calcium imaging in 3D intestinal organoids
Research into physiological intestinal processes such as nutrient absorption and drug interactions has traditionally relied heavily on animal experiments. This methodological publication presents a detailed protocol for live-cell calcium imaging in 3D intestinal organoids, providing a completely animal-free and human-specific alternative. Using the fluorescent dye Fura-2 AM, the protocol enables the dynamic and precise measurement of receptor activation and substrate transport in living cells. The 3D intestinal organoids demonstrate excellent dye uptake and reflect human intestinal physiology far more accurately than animal models or conventional 2D cell cultures. The method has broad applications, including the study of malabsorption syndromes, diarrheal diseases, and drug absorption across the intestinal epithelium. The publication demonstrates how complex metabolic and transport processes can be investigated directly in human-based models without involving animal suffering.