The symbiotic relationship between healthy hospital design and infection prevention and control (IPC) is crucial to developing a safe healthcare environment. Collaborative efforts in mitigating the risk of hospital-acquired infections (HAIs) are needed and will decrease morbidity, mortality rates, and costs. HAIs not only impact patient health but also tarnish the reputation of healthcare institutions. This paper delves into the distinctions between exogenous-derived and endogenous-derived HAIs, elucidating their sources, transmission mechanisms, and preventive strategies. Exogenous-derived HAIs can be prevented by a well-designed hospital layout which minimize contamination. Endogenous-derived HAIs originate from the patient’s own microbial flora, necessitating tailored infection prevention strategies such as antimicrobial prophylaxis. Standard precautions and transmission-based precautions, as outlined by the Centers for Disease Control and Prevention (CDC), form the cornerstone of infection control efforts. Hospital design should facilitate compliance with these measures, ensuring a microbial-safe environment conducive to patient recovery. Interdisciplinary collaboration between architects, healthcare professionals, and infection control experts are needed to integrate infection control principles into hospital design processes effectively. Key considerations include optimizing patient flows, separating clean and dirty materials, and implementing robust ventilation systems to mitigate airborne transmission risks. Furthermore, selecting appropriate surface materials resistant to microbial growth and enabling effective cleaning and disinfection protocols are important to maintain a microbial safe hospital environment. Most importantly, the shift towards single-occupancy rooms represents a significant stride in infection prevention, minimizing the risk of cross-contamination compared to multi-occupancy wards. Scientific evidence supports the efficacy of single-occupancy rooms in reducing microbial contamination and preventing HAIs.