Microencapsulation of bioactive compounds is a versatile and efficient strategy for promoting the stabilization and protection of labile, health-relevant molecules. Osmoporation is a promising technique for internalizing bioactive compounds utilizing yeast cells as a safe and efficient encapsulation system for multiple applications in the food and pharmaceutical industries. It is based on their natural osmoregulatory mechanisms that promote cell wall transient permeabilization as a response to sublethal osmotic perturbations. In this chapter, a detailed osmoporation protocol to produce yeast biocapsules is described, including the cultivation and harvest of food-grade yeast cells (Saccharomyces cerevisiae), preparation of osmo-modulatory solutions, and single- and multistage osmoporation procedures.