Being able to detect and respond to abiotic and biotic stresses is fundamental for plant growth and survival. However, understanding of signal transduction within the root remains limited. To help shed light on these processes, we have developed a bidirectional-dual‐flow‐RootChip (bi-dfRC), which adds bidirectional stimulation to the existing asymmetric laminar flow root perfusion platforms. In this paper we show design, fabrication and characterisation of the bi-dfRC, as well as growth of wildtype and Ca2+ indicator (G-CaMP3) Arabidopsis thaliana plants on the platform. Applicability of the bi-dfRC is further demonstrated by probing the dynamic response of Arabidopsis roots to simulated drought stress effects via a fluorescent Ca2+ sensor in a variety of combinations and spatial orientations. The latter enables the tracking of growth, localisation, and quantity in response to bidirectional stimulation in real time at a cellular level.