Fluorescence microscopy of etched methacrylate sections improves the study of mitosis in plant cells

Etched sections of methacrylate infiltrated plant tissue [Gubler (1989) Cell Biol. Int; Rep., 13:137-145; Baskin et al. (1992) Planta, 187:405-413] offer many advantages over the more traditional squash technique of Wick et al. [(1981) J. Cell Biol. 89:685-690] for immunofluorescence microscopic investigation of the plant cytoskeleton, especially during mitosis. These advantages include: (1) unimpeded access of antibody probes, (2) confocal-like imaging without the expense of confocal equipment, (3) maintenance of organ architecture as well as intracellular structure, (4) the ability to independently examine separate focal planes with the same or multiple antibody(s) or other labelling compounds, and (5) the ability to archive unetched sections, polymerized or non-polymerized infiltrated tissue. In this paper examples of staining of various microtubule cytoskeletal and mitotic proteins are shown in a variety of methacrylate embedded plant tissues.