Crop improvement, conservation and utilization of plant genetic resources has benefited from chromosomal studies of germplasms. In the Indian sub continent, lentil (Lens culinaris Medik.) is a major rabi pulse crop with 2n = 2x = 14 chromosomes. Chromosomal studies were carried out on Indian lentils during the period of 1952 to 1991 employing orcein squash techniques by different authors but there was no unanimity on chromosome morphology and chromatin length. This is the first study conducted on two wild and 12 cultivated cultivars of Lens for detailed and comparative chromosomal analysis using enzymatic maceration and air drying (EMA) based Giemsa staining methods. Chromosomal analysis revealed chromosomal stability, uniform karyotype formula (3m + 1m (sat) + 2sm + 1st), one pair of interstitial sat in either chromosome number 3 or 4 and interesting variations in total chromatin length (53.6-121.2 μm) in all the studied cultivars. More studies on other species and cultivars along with the application of flurochrome dyes for characterizing apparently similar chromosomes at the cultivar level may be very useful for future crop improvements.