[Objectives]  To investigate the tissue-specific expression of the male-specific gene Nilaparvata lugens NlEF7 and thereby provide a reference for multi-tissue dissection and in situ hybridization techniques for this species. [Methods]  Female and male adult N. lugens of different developmental stages were selected and dissected under a microscope to obtain salivary glands, intestines, female and male genitalia and other tissues. RNA was then extracted to analyze NlEF7expression in different tissues with quantitative PCR. After fixing samples with paraformaldehyde, RNA in situ hybridization detection was then performed using a digoxigenin-labeled NlEF7 probe and NBT/NBCT color development technology. [Results]  Detailed anatomical microscopic images of salivary glands, intestines and internal genitalia were obtained. The latter show morphological changes in the internal genitalia of female N. lugens that can confirm a female’s mating status. The results of quantitative PCR and in situ hybridization indicate that the NlEF7 gene was specifically expressed in the internal genitalia and fat body of males, and that expression was highest in the testis. There was almost no hybridization signal after fixation and in situ hybridization of the intact body. Destructive treatment of insects, or separation of tissue samples, followed by fixation and hybridization can help increase the intensity of the hybridization signal. [Conclusion]  The NlEF7 gene is specifically expressed in the internal genitalia and fat body of adult male N. lugens.