In this paper, we present in detail the design, both electromagnetic and mechanical, the fabrication, and the test of the first prototype of a Scanning Microwave Microscope (SMM) suitable for a two-port transmission measurement, recording, and processing the high frequency transmission scattering parameter S 21 passing through the investigated sample. The S 21 toolbox is composed by a microwave emitter, placed below the sample, which excites an electromagnetic wave passing through the sample under test, and is collected by the cantilever used as the detector, electrically matched for high frequency measurements. This prototype enhances the actual capability of the instrument for a sub-surface imaging at the nanoscale. Moreover, it allows the study of the electromagnetic properties of the material under test obtained through the measurement of the reflection (S 11) and transmission (S 21) parameters at the …