Self-assembly of non-ionic amphiphilic architectures into nanostructures with defined size, shape and morphology has garnered substantial momentum in the recent years due to their extensive applications in biomedicine. The manifestation of a wide range of morphologies such as micelles, vesicles, fibers, tubes, and toroids is thought to be related to the structure of amphiphilic architectures, in particular, the choice of the hydrophilic and hydrophobic parts. In this review, we look at different types of non-ionic small amphiphilic architectures and the factors that influence their self-assembly into various nanostructures in aqueous medium. In particular, we focus on the explored structural parameters that guide the formation of various nanostructures, and the ways these structures can be used in applications ranging from drug delivery to cell imaging.