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, 187:4 05-413] offer many advantages over the more traditional squash techniq ue of Wick et al. [(1981) J. Cell Biol. 89:685-690] for immunofluoresc ence microscopic investigation of the plant cytoskeleton, especially d uring mitosis. These advantages include: (1) unimpeded access of antib ody probes, (2) confocal-like imaging without the expense of confocal equipment, (3) maintenance of organ architecture as well as intracellu lar structure, (4) the ability to independently examine separate focal planes with the same or multiple antibody(s) or other labelling compo unds, and (5) the ability to archive unetched sections, polymerized or non-polymerized infiltrated tissue. In this paper examples of stainin g of various microtubule cytoskeletal and mitotic proteins are shown i n a variety of methacrylate embedded plant tissues. (C) 1998 Wiley-Lis s, Inc.