A wide variety of substances that may affect the development of the central nervous system (CNS) in human exist in environment. Evaluating how chemical teratogens adversely affect the developing brain may provide important insights not only for the management of industrial chemicals but also for the prevention and treatment of CNS diseases. However, a system for efficiently and comprehensively evaluating the developmental neurotoxicity (DNT) of chemicals has not yet been sufficiently established. Here, we try to establish the assessment of DNT analyzing the dynamics of neurons, astrocytes, and microglia in developing zebrafish CNS, with particular focus on the property of microglia reflecting neuroinflammation. We generated a transgenic zebrafish line expressing different fluorescent proteins; Cerulean for neurons, mCherry for astrocytes, and mVenus for microglia. Subsequently, we exposed various neurotoxicants to zebrafish embryo and performed in vivo fluorescence imaging. We are trying to quantitatively measure the total amount, distribution, and morphology of neurons, astrocytes, and microglia in the larval zebrafish brain by imaging analysis and detect DNT comparing with already reported findings.