Tobramycin is widely used in the treatment of bacterial infections, particularly those caused by Gram-negative bacteria. It is commonly administered as an injectable solution for severe infections such as sepsis, pneumonia, and infections of the urinary tract, abdomen, and skin. In ophthalmology, it is formulated as eye drops or ointments to treat bacterial conjunctivitis and other eye infections.

In cystic fibrosis patients, tobramycin is often used in inhaled form to manage chronic Pseudomonas aeruginosa infections in the lungs, helping to reduce bacterial load and improve respiratory function. Its effectiveness against resistant strains of bacteria makes it a critical component in combating hospital-acquired infections. Additionally, it is sometimes used in combination with other antibiotics to enhance its antibacterial spectrum and efficacy.