The current study aims to produce stable liposomes from total lipid extracts from bacteria. Liposomes are the small vesicles that are made up of lipids. On their structural basis, they can be considered as simplified cell structure of cell membrane. Structure of liposomes depends on the pH of preparation buffer, method of preparation and the environmental condition in which they are prepared. Liposomes have importance in the field of medicines for diagnostic and therapeutic purposes. They mainly work as a vehicle for drug delivery. The objective of the current study was to make stable liposomes from two types of bacterial samples i.e., a Gram-positive and a Gram-negative strain. E. coli and Bacillus sp. were selected as representative of Gram-negative and Gram-positive bacteria, respectively. Lipid extraction was performed by various methods, out of which the modified Bligh and Dyer method gave most effective results. Liposomes were prepared by extrusion and their stability and efficiency was tested by fluorescence spectrophotometer using OxanolVI. Our results showed that liposomes formed by lipids extracted from E. coli were more stable than the liposomes formed by lipids extracted from Bacillus sp. Keywords: liposomes, Lipids, diagnostic, therapeutic, purposes, fluorescence, Bacillus sp.