The genus Nymphoides (Menyanthaceae), commonly known as “floating hearts,” is one of the most ubiquitous floating leaved genera belonging to the family Menyanthaceae having about 50 species widely distributed worldwide in tropical and temperate regions. This aquatic genus is economically valuable as decorative plants, vegetables, and a common medication in Ayurvedic medicine due to its anticonvulsant, antioxidant, hepatoprotective, cytotoxic, and anticancer properties. Pollen cryopreservation offers a unique and efficient method for long-term storage of viable pollen, crucial for the conservation and utilization of economically and medicinally important plant species like Nymphoides. In this study, standardized protocols were developed for three Nymphoides species (N. indica, N. geminata, and N. hydrophyllum), encompassing pollen collection, dehydration, in vitro germination, and cryopreservation in liquid nitrogen. These protocols ensure the preservation of genetic resources and support further studies on the reproductive biology and conservation of this valuable aquatic genus. The details of material and methods used in development of protocol are discussed in this chapter.