Secure quantum communication systems (QCS) based on the transmission of crucial information through single photons are among the most appealing frontiers for telecommunications, though their development is still hindered by the lack of cheap and bright single photon sources (SPSs) operating at room temperature (RT). In this paper, we show the occurrence of photon antibunching at RT from single colloidal CdSe/ZnS nanocrystals (NCs) inserted in a vertical microcavity. Moreover, by using high-resolution lithographic techniques, we conceived a general route for positioning single colloidal quantum dots in the microcavity. The findings and the technique presented here can be considered a first step towards the development of SPS devices operating at RT.