This chapter presents in vitro protocols for studying interactions between Cryptococcus neoformans and microglial cells. These phagocytes are a critical component of the central nervous system's immune response to C. neoformans, the most common fungal agent of meningoencephalitis worldwide. Given its severity and high incidence in immunocompromised patients, understanding the interaction of C. neoformans with central nervous system (CNS) cells could help advance treatment strategies. These protocols leverage microscopy and molecular biology techniques to elucidate the mechanisms of C. neoformans invasion and persistence within the CNS, offering a methodological framework for fungal neuropathogenesis research.