Abstract
High-pressure freezing is a method of specimen preparation for electron microscopy that freezes samples without significant ice crystal damage. It is effective for preparation of electron microscopic samples in plants and has advantage to conserve antigenicity in immunolabeling. We have improved a combination method of high-pressure freezing and freeze substitution for plants using a high-pressure freezing machine, EM PACT (Leica). To respond promptly to needs from researchers both inside and outside RIKEN Plant science center, we aim to construct an electron microscopic sample library of model plants. Until now, we have prepared samples of Arabidopsis, Tobacco, Tobacco cultured cell BY-2, Arabidopsis cultured cell and rice cultured cell. Each sample was substituted by three types fixative, in order to apply various antigens and antibodies. Moreover, we are improving cryoultramicrotomy for plant cells and tissues.
