title: Immunoelectron Microscopy of Cryofixed and Freeze-Substituted Plant Tissues.
abstract: Cryofixation and freeze-substitution techniques provide excellent preservation of plant ultrastructure. The advantage of cryofixation is not only in structural preservation, as seen in the smooth plasma membrane, but also in the speed in arresting cell activity. Immunoelectron microscopy reveals the subcellular localization of molecules within cells. Immunolabeling in combination with cryofixation and freeze-substitution techniques provides more detailed information on the immunoelectron-microscopic localization of molecules in the plant cell than can be obtained from chemically fixed tissues. Here, we introduce methods for immunoelectron microscopy of cryofixed and freeze-substituted plant tissues.
authors: Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyou-ku, Tokyo, 113-8657, Japan.
authors: Miyuki
authors: M
authors: Takeuchi
authors: Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan.
authors: Keiji
authors: K
authors: Takabe
authors: Department of Picobiology, Graduate School of Life Science, University of Hyogo, 2167 Shosha, Himeji, Hyogo, 671-2280, Japan. mineyuki@sci.u-hyogo.ac.jp.
authors: Yoshinobu
authors: Y
authors: Mineyuki
mesh_terms: D000906:Antibodies / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D015925:Cryopreservation / Q000379:methods*; D004853:Epoxy Resins / Q000737:chemistry; D005404:Fixatives / Q000737:chemistry; D017110:Freeze Substitution / Q000379:methods*; D015870:Gene Expression; D005976:Glutaral / Q000737:chemistry; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods; D008867:Microtomy; D019697:Onions / Q000378:metabolism / Q000648:ultrastructure*; D059828:Plant Cells / Q000378:metabolism / Q000648:ultrastructure*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000378:metabolism / Q000648:ultrastructure; D032107:Populus / Q000378:metabolism / Q000648:ultrastructure; D012639:Seeds / Q000378:metabolism / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods; D016610:Tissue Embedding / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D000906:Antibodies; D004853:Epoxy Resins; D005404:Fixatives; D018398:Homeodomain Proteins; D010940:Plant Proteins; D014404:Tubulin; D005976:Glutaral
keywords: Cryofixation; Freeze substitution; High-pressure freezing; Immunoelectron microscopy; Plant tissue