Aphids are serious agricultural pests capable of making direct damages to crops and of transmitting hundreds of species of pathogens that cause considerable losses worldwide. Their piercing-sucking mouthparts are well-adapted structures to pierce plant tissues and uptake sap and nutrients from their host plants. They consist of four long needle-like cuticular stylets synthesized at each molt by specialized glands. Our previous work has shown that proteins are not evenly distributed in the stylets conferring local functionalization of cuticular micro-territories such as the acrostyle with stylin proteins emerging at the stylet surface involved in virus binding. To better understand when and how the acrostyle is synthesized, we developed several microscopy and high resolution imaging approaches, such as X-ray synchrotron-based micro computed tomography, or multiplex RNA imaging on whole-mount cleared aphids. Our results provide an unprecedented level of detail on the stylet biogenesis in the aphid species Acyrthosiphon pisum.