This paper explores the making of modern medical knowledge and practice in the late Ottoman Empire by focusing on the role of the Bakteriolojihane-i Hümayun (Royal Bacteriology Institute) which was established in 1894 as part of a broader scientific and diplomatic effort to control the devastating impact of the 1892 cholera epidemic in Istanbul. Maurice Nicolle, who was in charge of microbiology at the Pasteur Institute in Paris, served as the head of the Bakteriolojihane until his resignation in 1901. Literature on the Bakteriolojihane focuses on Nicolle’s tough personality and conflictual relationships with those in the political and medical circles. Decentering the emphasis on Nicolle’s persona, this paper analyzes imperial and local dynamics that formed medical education as well as knowledge production and practices that took place in the Bakteriolojihane. The establishment of the Bakteriolojihane overlapped with a particular historical context highlighted by the intensification of scientific competition between followers of Pasteur and those of Koch over the dissemination of research and methods on bacteriology. This paper examines the Bakteriolojihane and the world around it by locating the institute in this competitive framework and addressing questions regarding the relationship between the nature of sovereignty and the production of scientific knowledge in the late Ottoman context. While it had been a common practice among the medical-bureaucratic circles in the Ottoman Empire to invite European medical experts as advisers, the Bakteriolojihane constituted the first institutional example of such practices in this historical context. I will explore the notion of “colonial medicine” by looking at the interplay between medical knowledge and practices in the Bakteriolojihane at a time of scientific colonial competition. Medical institutions like the Bakteriolojihane also functioned as schools to educate medical personnel who later played significant roles in the medical-bureaucratic positions. The number of medical students who visited schools and hospitals in Europe, predominantly in France, for advanced study increased sharply after the establishment of the Bakteriolojihane. By focusing on the experiences of these medical experts in multiple contexts, this paper addresses questions concerning the political identities, social perceptions and medical expertise of these physicians in the framework of their encounter with the European medical setting as well as the ways in which they translated their experiences into domestic conditions, especially in the Anatolian countryside, as they became policy makers and medical practitioners.