Meiosis halves the number of chromosomes to produce haploid gametes through nuclear division twice following single round of DNA replication, and is essential for eukaryotic sexual reproduction. Arabidopsis thaliana is a traditional model organism used for molecular genetic study. Recently, with breakthroughs in microscopic technologies for observing chromosomes, analyses of chromosome morphology during meiosis using cytological methods have made great advances to understand molecular and genetic mechanisms in regulation of meiosis. In this study, we described in detail the chromosome spread technique we developed, which is well used for observation of chromosome behaviors in Arabidopsis male meiocytes.