This study investigated the impact of six different doneness temperatures (rare, medium-rare, me- dium, medium-well, well-done, and very well-done) on the sensory qualities, physicochemical properties, and microbiological characteristics of indigenous beef from Bangladesh. The beef was cooked at the following temperatures: Rare: 52 °C, Medium-rare: 57 °C, Medium: 65 °C, Medium- well: 66 °C, Well-done: 71 °C, Very Well-done: 80 °C. The results demonstrated that the sensory qualities of the meat varied dramatically with cooking temperature. Rare meat had the most soft and juicy texture, whereas, extremely well-done beef had the most stiff and dry feel. The physico- chemical parameters of the beef, such as moisture content, protein content, and fat content, also varied dramatically with cooking temperature. Rare meat had the highest moisture content and lowest fat level, while extremely well-done beef had the lowest moisture content and highest fat content. The microbiological properties of the beef also changed dramatically with cooking tem- perature. Rare beef had the highest microbial burden, while extremely well-done beef had the low- est microbial load. This is because cooking at higher temperatures kills more bacteria. Previous Research has explored the effects of varying doneness temperatures on the sensory, physicochemi- cal, and microbiological aspects of beef. However, these investigations have been conducted on beef from different nations and have employed different cooking methods. Therefore, there is a research void on the impacts of doneness temperature on the sensory, physicochemical, and micro- biological features of indigenous beef of Bangladesh. The outcomes of this study have significance for optimizing cooking procedures to increase the sensory appeal and ensure the microbiological safety of indigenous beef. The data also imply that well-done meat may be a healthier alternative than medium-rare or medium steak, as it has a reduced microbial load. However, it is crucial to remember that Very Well-done beef should be avoided, as it has a dry and harsh texture. The out- comes of this study revealed that the doneness temperature of well-done, which is 71℃, fits and satisfies the consumers' unique preferences and health concerns.