This article examines tensions that professionals in healthcare settings in low-to-middle income countries (LMICs) face in the evolving field around surveillance of antimicrobial resistance (AMR). Few public health problems are of greater global importance today than AMR, that poses a threat to our ability to treat infections. In this context, the microbiology laboratory occupies a prominent place and the knowledge field of microbiology is expanding. In this study, we interviewed twenty-three (n = 23) professionals with expertise on AMR and public health systems to synthesise knowledge on strengthening AMR surveillance in LMICs. By drawing on a practice approach [Schatzki, T. R. 2001. "Practice Theory." In "The Practice Turn in Contemporary Theory" (1-14), edited by T. R. Schatzki, K. Knorr-Cetima, and E. von Savigny. New York: Routledge.] combined with socio-cultural and cultural-historical theories (CHAT) [Engeström, Y. 1987. "Learning by Expanding: An Activity-Theoretical Approach to Developmental Research." Helsinki: Orienta-Konsultit] the analysis reveals seven tensions between elements of the systems and discusses how such tensions serve to frame implications for implementing a capacity strengthening programme. The analysis shows that the novelty of the AMR as well as being a multi-disease and multisectoral by nature challenges existing forms of professional practice in healthcare settings. It also suggests that AMR requires to be dealt with through inter-professional and inter-sectoral approaches, while maintaining a focus across the local, national, and global systems, which is essential for initiatives that are set to address challenges to global health.