[Aim]  To examine how different female colorations, along with surface substances on the cuticle of the ladybird Harmonia axyridis, before and after death, affect male mating preferences. [Methods]  A preliminary analysis of the composition of the main cuticle surface substances of males and females, and live and dead, H. axyridis was conducted. Behavioral analysis software and simulated ladybirds were then used to investigate differences in behavior towards various potential mates. Solvent extraction and GC-MS analysis were carried out on elytra from several different colorations of live and dead female H. axyridis, to qualitatively identify surface substances, and quantitatively analyze the differences between different colored, and between live and dead, individuals. [Results]  Male H. axyridis exhibit a mating preference for females with black elytra; however, the attractiveness of elytra washed with hexane to males was significantly reduced (P < 0.01). Males were able to identify dead individuals during the mating process. Chemical analysis confirmed significant differences (P < 0.05) in two hydrocarbons—9-nonadecene and 1,15-pentadecanediol—between sexes and across elytra with different background colors. One week after the death of H. axyridis, significant quantitative changes were detected in four surface substances on the elytra, but these compounds could not be successfully identified. [Conclusion]  The surface substances of female H. axyridis elytra influence male mating preferences, with different colored females varying in their level of attractiveness towards males. Recognition of dead females may also be linked to differences in elytra compounds. Systematic research on the surface substances of ladybirds both improves our understanding of their mating processes and the artificial breeding of these useful biological control agents.