Epilepsy is a clinical syndrome caused by highly synchronized abnormal discharges of neurons in the brain due to a variety of causes. The clinical manifestations are characterized by seizures that are transient, repetitive and stereotyped. Lipid droplets (LDs) are cellular organelles that help to maintain cellular functioning. Persistent epilepsy is associated with altered metabolism of LDs. Hesperidin as a natural product used in treatments related to neurological disorders. Therefore, studying the effect of hesperidin on cellular LDs during epilepsy has become a priority. Based on this problem, the near-infrared probe YCN was used to evaluate the effect of hesperidin on epileptic cells. First, the probe YCN had good photostability, biocompatibility and excellent targeting ability for LDs. YCN could detect cellular changes induced by honokiol, plumbagin or hesperidin, demonstrating the ability of the YCN probe to sensitively detect changes in the intracellular environment. Most importantly, YCN was used to image changes in epileptic cells in situ and to explore the effects of hesperidin on epileptic cells, revealing the significant antiepileptic activity of hesperidin at certain concentrations. The effective application of the probe YCN could aid the development and efficacy evaluation of antiepileptic drugs.