Source: LeSavage, B. L., et al. Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostaining of Cells in 3D. J. Vis. Exp. (2018).
This video demonstrates a technique for immunostaining cells encapsulated in a 3D hydrogel. The hydrogel culture containing neural progenitor cells is fixed, permeabilized, and treated with a blocking solution to prevent non-specific immunostaining. A primary antibody cocktail is added to bind specific proteins, followed by fluorescently labeled secondary antibodies and a nuclear stain. The sample is then mounted on a slide and observed under a confocal microscope.