Poor adherence to oral antiretroviral therapy (ART) remains an important challenge in the treatment of HIV. Microneedles (MN) potentially could offer a non-invasive long-acting (LA) delivery approach, avoiding the need for daily dosing of ART. However, this claim has yet to be explored amongst its potential end-users. The aim of this mixed methods study was to investigate the perspectives from various end-users surrounding the translation of MN technology to general clinical practice, with a particular focus on delivery of ART. Quantitative postal questionnaires were distributed amongst healthcare professionals (HCPs) and the lay public (LP). A total of 208 responses were obtained (HCP, 69; LP, 139), with a completion rate of 34.7%. The consensus on MN technology was positive from both demographics (HCP, 97.1%; LP, 98.6%), with further strong support of postulated MN use within HIV (HCP, 97.1%; LP, 98.6%). Qualitative focus groups were employed to investigate in-depth, the perspectives of 12 patients with HIV. Again, consensus on MN technology was positive, highlighting benefits pertinent to HIV, including discreet self-application and potential sustained release thus avoiding daily oral ART and associated side effects. Patient concerns focused on the need for varied MN dosing schedules and a reluctance to change from established ART. The findings of this study provide an initial indication of MN acceptability, particularly for use within HIV, from various end-user demographics. Furthermore, concerns raised advocate the importance of continued translational research in this area and should act as motivators for those in MN development to ensure a patient-centred MN product is delivered. Graphical Abstract.