This chapter explores the concept of karyotype and its analysis, a fundamental tool in understanding the genomic architecture of various species. Introduced by Russian scientist Levitsky in 1931, the term “karyotype” refers to the unique chromosome complement of each species, represented visually through cytogenetic methods. The relevance of karyotype analysis in the genomic era lies in its efficiency for visualizing critical genomic information such as chromosome number, morphology, and banding patterns. Karyotyping remains unaffected by environmental or developmental variables, thus it offers consistent insights into chromosomal characteristics. Plant karyotyping, however, presents distinct challenges due to factors like cell wall presence and polyploidy. This chapter addresses the objectives of karyotype analysis, including assessing chromosome features crucial for cytotaxonomy and plant breeding, and identifying chromosomal abnormalities. It also discusses karyotype symmetry and asymmetry, emphasizing size variability and its evolutionary implications, as proposed by Stebbins in his qualitative-quantitative framework. The classification into symmetric and asymmetric karyotypes further elucidates chromosomal organization, with symmetric karyotypes signifying a more primitive genomic state. Modern advancements in cytogenetic techniques, such as fluorescence in situ hybridization (FISH), have enhanced the understanding of plant genetic complexity, paving the way for more sophisticated research in karyotype analysis.