Bacterial biofilms are aggregations of bacterial cells embedded into self-produced matrix of extracellular polymeric substrates (EPS). Biofilms develop on or within indwelling medical devices which eventually lead to nosocomial infections. Here we describe the development of a PDMS-based flow cell device which mimics catheters used in hospitals. The device was fabricated using soft lithography technique. Staphylococcus aureus and Acinetobacter baumanii which are known to form biofilm and are associated with urinary tract infections were used as representative organism's to demonstrate biofilm formation in the device. Quantitative assay of biofilm formed was done using Microscopy (Z-stacking). This device can be successfully used for semi-quantitative detection of bacterial biofilms and is ideally suited for use in screening of anti-biofilm compounds.