Pseudomonas aeruginosa is a leading cause of disease and death particularly in cystic fibrosis patients and also considered resistance to chemotherapeutic agents. Therefore, it is very difficult to remove the Pseudomonas aeruginosa from the hospital environment by using simple techniques. In the contemporary study, biofilm mediated mechanism of various antimicrobial responses were analyzed. For this purpose, different Pseudomonas aeruginosa clinical isolates were collected from Pakistan medical institute Islamabad (PIMS) hospital and were investigated for pellicle formation. Pseudomonas aeruginosa isolates were studied for different groups of antibiotics including imipenem, meropenem, ceftazidime, amikacin, tobramycin, gentamicin, piperacillin, cefoperazone, and cefotaxime. The goal was to check antimicrobial susceptibility of pseudomonas aeruginosa which shows resistant to tobramycin, imipenem, meropenem, amikacin, gentamicin, cefotaxime, piperacillin, ceftazidime, cefoperazone. Additionally, in this study, Pseudomonas aeruginosa strains were also investigated for pellicle formation. In conclusion, this research work wills highlights the useful mechanism of antibiotics resistance to Pseudomonas aeruginosa infections in clinical practice. Keywords: Antibiotics, Pseudomonas aeruginosa, antibiotics, Biofilm, Peliclle.