As endogenous chemical substances, neurotransmitters play a vital role in maintaining normal life activities. Abnormal levels of neurotransmitters can lead to physical, mental, and some neurodegenerative diseases. However, the ultralow concentration, complex chemical properties, and release modes of neurotransmitters make their accurate detection in vivo a great challenge. To accurately monitor neurotransmitters in the brain and accurately understand the release kinetics of neurotransmitters, we reviewed several method commonly used in the past five years to detect neurotransmitters in vivo and their research progress. The basic principle and applicability of microdialysis, electrochemical sensors, and fluorescence sensors are introduced in detail.