Nitric oxide (NO) acts as a signalling molecule in the brain. NO has been implicated in a variety of central functions such as learning, plasticity and neurodegeneration. It is also involved in regulation of autonomic homeostasis at different levels of neuraxis including the nucleus tractus solitarii. In spite of the ample evidence for NO-mediated signalling many aspects of its mechanism of action the brain remain unknown largely due to the difficulties of NO detection in real time coupled with its unique ability to freely cross cellular membranes. Here we give a brief overview of the currently available options for NO detection in the brain (such as electrochemistry, fluorescent indicators, electron-paramagnetic resonance) and consider some of their limitations. We conclude that it would be extremely useful to develop a highly sensitive probe for NO detection with some kind of build-in amplification which would magnify the changes triggered by NO to allow its detection within microdomains of the brain tissue in real time.