Fluorescence in Situ Hybridization (fish) on Vibratome Sections of Plant Tissues

This protocol describes the application of fluorescence in situ hybridization (FISH) on three-dimensionally preserved tissue sections derived from intact plant structures such as roots or florets. The method is based on the combination of vibratome sectioning with confocal microscopy. The protocol provides an excellent tool to investigate chromosome organization in plant nuclei in all cell types and has been used on tissues of several different plant species. The visualization of three-dimensionally wellpreserved tissues means that cell types can be confidently identified. For example meiocytes can be clearly identified at all stages of meiosis and can be imaged in the context of their surrounding maternal tissue. FISH can be used to localize centromeres, telomeres and sub-telomeric regions, and total genomic DNA can be used as probes to visualize introgressed chromosomes or chromosome segments.