Abstract
The protocol provides fully detailed steps for preparing microscopic slides of acrylamide-embedded maize meiotic cells. This method is particularly useful for examining chromatin structure and chromosome arrangement without destroying the three-dimensional organization of the nucleus.
Keywords: Three-dimensional microscopy, Immunolocalization, Meiocytes, Chromosomes, Maize
Background
Meiosis is a dynamic process involving homologous chromosomes pairing, synapsis, and recombination. Cytological analysis to investigate localization and dynamics of meiotic proteins are crucial for understanding details of these processes. In many microscopic slide preparation methods, spatial chromatin organization is damaged by either mechanical processing or chemical solvents. Here, we describe a three-dimensional microscopy protocol to analyze chromatin structure and meiotic protein localization without disturbing nuclear organization.
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
We thank Mateusz Zelkowski for pictures and video. Research to improve this protocol was supported by a grant from the National Science Foundation (IOS-1848788). This protocol is an adaptation of procedures of Franklin et al. (1999) and Ronceret et al. (2009). Authors declare no competing interests.
References
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