Sexual reproduction requires creating haploid gametes from diploid parents through meiosis. Prior to meiotic cell divisions, homologous chromosomes pair and recombine to form the meiotic bivalent structure in prophase. Telomeres are attached to the nuclear envelope, and the cytoskeleton network outside the nucleus drives the chromosome movement to efficiently pair homologous chromosomes. Meiotic prophase is followed by meiosis I division, where homologous chromosomes segregate away to reduce the ploidy (reductional division). In the following meiosis II division, sister chromatids segregate in a manner similar to mitosis (equational division). Chromosome segregation in both meiotic divisions is mediated by the interaction between centromeres and the spindle machinery. Therefore, it is critical to investigate chromosome and cytoskeleton dynamics during both meiotic prophase and meiotic divisions to understand (1) how chromosomes find their homologous partners, (2) how faithful chromosome segregation is achieved, and (3) how these processes can go wrong or exploited by selfish DNA, causing fertility issues. Recent advances in technology provided new angles to tackle these open questions in the meiosis field. This Methods Collection covers newly-developed techniques to investigate both chromosome and cytoskeleton dynamics in meiosis, using different species.