This chapter delves into the process of gametic chromosome analysis in diploid higher plants, focusing on the role of flowers as the primary source of haploid gametes required for sexual reproduction. It outlines the meiotic division occurring in the anthers, where pollen mother cells (PMCs) generate haploid daughter nuclei. The chapter emphasizes the importance of using fresh flower buds to prepare meiotic chromosomes, highlighting Barbara McClintock’s method as a notable advancement. The preparation techniques, notably the conventional carmine smear method, are detailed, providing a step-by-step approach for creating cytological slides. The process requires specific materials, such as flower buds, slides, and aceto-carmine solution, as well as instruments such as needles and scalpels. The meticulous collection and preparation of flower buds, removal of anthers, and the application of staining solutions are outlined to achieve clear visualization of the meiotic chromosomes. It also touches upon the variance in bivalent numbers and the physical characteristics of haploid cells. Through documented case studies of various plant materials, the chapter serves as a practical guide for undergraduate and postgraduate students in understanding meiotic chromosome preparations. The methodologies enabled an efficient observation of different meiotic stages, enhancing comprehension of gametic chromosome analysis. Ultimately, the chapter highlights the crucial elements for successful gametic chromosome studies, contributing to the broader understanding of plant reproductive biology and genetics.