Cryptococcus neoformans is a pathogenic yeast characterized by the production of a conspicuous polysaccharide capsule, which is critical for its virulence and ability to evade the host immune response. The capsule, a complex structure composed of 88% glucuronoxylomannan (GXM) and 12% galactoxylomannan (GalXM) surrounds the fungal cell and plays an important role in pathogenicity. A special feature of the C. neoformans capsule is its hydrophilicity and the fact that its refractive index is very close to that of the surrounding medium, which makes it invisible under the light microscope without special techniques. Capsule formation is a dynamic process influenced by growth conditions and can be visualized using negative staining methods, the most commonly used method being India ink staining. In this protocol, we describe the standard techniques for triggering capsule formation and subsequent visualization of the capsule using India ink negative staining in two different growth media: Dulbecco’s Modified Eagle Medium (DMEM) and Sabouraud Dextrose Broth (SDB). DMEM is often used for co-culture experiments with mammalian cells and provides conditions more similar to those in a host environment. SDB, on the other hand, is often used for fungal cultures and supports the robust growth of C. neoformans. Capsules can be effectively induced in both DMEM and SDB, although size and structure may vary depending on the medium used. After incubation, India ink is added to the fungal suspension to stain the background while the capsule remains unstained. This creates a clear “halo” around each cell, which is characteristic of capsule formation (Gutierrez-Gongora et al, Sci Rep 13:4928, 2023). This method allows a qualitative analysis of the capsule size, shape and overall capsule density of the fungal cells. The capsule is a key factor in studies of virulence, immune evasion and pathogenesis, and this protocol serves as an important tool to visualize and quantify the capsule in laboratory settings. This method allows researchers to study the influence of different environmental conditions, antifungal treatments and genetic factors on the production of the C. neoformans capsule, which is an important aspect of its pathogenic potential (Zaragoza et al, Adv Appl Microbiol 68:133–216, 2009).