Background:Breast cancer has emerged as a leading site of cancer among women in India. Fine needle aspiration cytology (FNAC) has been routinely applied in assessment of breast lesions. Cytological evaluation in breast lesions is subjective with a “gray zone” of 6.9–20%. Quantitative evaluation of nuclear size, shape, texture, and density parameters by morphometry can be of diagnostic help in breast tumor.Aims:To apply nuclear morphometry on cytological breast aspirates and assess its role in differentiating between benign and malignant breast lesions with derivation of suitable cut-off values between the two groups.Settings and Designs:The present study was a descriptive cross-sectional hospital-based study of nuclear morphometric parameters of benign and malignant cases.Materials and Methods:The study included 50 benign breast disease (BBD), 8 atypical ductal hyperplasia (ADH), and 64 carcinoma cases. Image analysis was performed on Papanicolaou-stained FNAC slides by Nikon Imaging Software (NIS)–Elements Advanced Research software (Version 4.00). Nuclear morphometric parameters analyzed included 5 nuclear size, 2 shape, 4 texture, and 2 density parameters.Results:Nuclear morphometry could differentiate between benign and malignant aspirates with a gradually increasing nuclear size parameters from BBD to ADH to carcinoma. Cut-off values of 31.93 μm2, 6.325 μm, 5.865 μm, 7.855 μm, and 21.55 μm for mean nuclear area, equivalent diameter, minimum feret, maximum ferret, and perimeter, respectively, were derived between benign and malignant cases, which could correctly classify 7 out of 8 ADH cases.Conclusion:Nuclear morphometry is a highly objective tool that could be used to supplement FNAC in differentiating benign from malignant lesions, with an important role in cases with diagnostic dilemma.