The nuclear membrane also referred as nuclear envelope (NE) encloses the nucleoplasm of the cell and acts as a barrier between the nuclear DNA and the cytoplasm. Recent studies suggest that the composition of proteins at the NE is not even throughout the whole membrane and can show zones with accumulation of some proteins inner and outer nuclear membrane protein, emerin, lamin A/C, or nesprin-2 in relation to cells’ polarization. Here we propose a methodology that combines immunofluorescent staining with quantitative imaging to prepare maps of protein distribution frequency that can serve to compare protein distribution on nuclear membrane.