The excessive glutamate-induced neuroexcitotoxicity has been widely thought as the main cause of brain cell damage in severe neurological diseases (such as ischemic stroke). Electrochemical technology is the most commonly used tool for glutamate monitoring at present. It could be applied to provide rapid and accurate assessment of the neuroexcitotoxicity for patients and animal disease models. Research advances in glutamate sensors will be important in establishing efficient therapies and brain protection strategy, and developing new drugs for severe neurological diseases. Application of nanotechnology, new enzymes, photolithography, printing, transistor technology in fabrication of glutamate sensors have greatly boosted developments in electrochemical monitoring of glutamate. In the present paper, research progress of sensing principles, design and preparation technology as well as the applications in medical studies have been reviewed, and future trends and prospective have also been discussed.