A self-adhesive layered septum is disclosed. In one example, the septum includes a first outer layer including a thermoplastic elastomer such as a styrenic block copolymer, containing styrene ethylene butylene styrene (TPE-SEBS) capable of closing at least partially an aperture formed when a needle is inserted through the layer; an adhesive second layer for adhering the septum to a mouth area of a well or container to which the septum is attachable; and a thermoplastic third layer between the first and second layers, thermobonded to the first layer and providing better adherence for the adhesive layer. The first layer includes a recess and a vent, which reduce pressure differentials in use, but together with the third layer minimize evaporation through the septum.