Planing frozen hydrated plant specimens for SEM observation and EDX microanalysis Export

@article{citeulike:2058711, abstract = {A procedure is described for forming a flat face on a frozen piece of plant tissue, which may then be observed fully-hydrated or lightly etched, and coated or uncoated with a metal film, in scanning electron microscopy (SEM). The frozen sample was planed with a glass knife at -80°C in cryo-ultramicrotome. The sections were discarded, and the planed block face placed on the cold stage in the microscope column, either for observation uncoated at low kV, or for light etching (-90°C) to reveal the cell outlines. If a higher accelerating voltage was needed, the face was given an evaporative coating of Al in the cryo-preparation chamber and returned to the column. The advantages of the planed face over the usual fracture face are illustrated: imaging at a chosen rather than a chance position; clearer cellular and subcellular detail; preservation of hydrated gels like mucilage and swollen cell walls; the possibility of making serial parallel sections through the same piece of tissue; opportunities for accurate morphometric analyses on the planed face; capacity to produce longitudinal sections; preservation of very delicate structures that are destroyed by fixation and drying. A major advantage of the Al-coated planed face is the increased accuracy of energy-dispersive X-ray (EDX) microanalyses on a smooth rather than a rough surface. Tests are included which show that neither the light etching employed, nor successive planing, interferes with the analyses of elements in the frozen face. {\copyright} 1994 Wiley-Liss, Inc.}, address = {Cryo-Analytical SEM Facility, Carleton University, Ottawa, Canada K1S 5B6; Biology Department, Carleton University, Ottawa, Canada K1S 5B6; Department of Biological Sciences, Lancaster University, Lancaster LA1 4YQ, England}, author = {Huang, Cheng X. and Canny, Martin J. and Oates, Kenneth and Mccully, Margaret E.}, citeulike-article-id = {2058711}, citeulike-linkout-0 = {http://dx.doi.org/10.1002/jemt.1070280108}, citeulike-linkout-1 = {http://www3.interscience.wiley.com/cgi-bin/abstract/109897690/ABSTRACT}, doi = {10.1002/jemt.1070280108}, journal = {Microscopy Research and Technique}, keywords = {cryo, techniques}, number = {1}, pages = {67--74}, posted-at = {2007-12-05 03:28:39}, priority = {2}, title = {Planing frozen hydrated plant specimens for SEM observation and EDX microanalysis}, url = {http://dx.doi.org/10.1002/jemt.1070280108}, volume = {28}, year = {1994} }

TY - JOUR ID - citeulike:2058711 L3 - citeulike-article-id:2058711 N2 - A procedure is described for forming a flat face on a frozen piece of plant tissue, which may then be observed fully-hydrated or lightly etched, and coated or uncoated with a metal film, in scanning electron microscopy (SEM). The frozen sample was planed with a glass knife at -80°C in cryo-ultramicrotome. The sections were discarded, and the planed block face placed on the cold stage in the microscope column, either for observation uncoated at low kV, or for light etching (-90°C) to reveal the cell outlines. If a higher accelerating voltage was needed, the face was given an evaporative coating of Al in the cryo-preparation chamber and returned to the column. The advantages of the planed face over the usual fracture face are illustrated: imaging at a chosen rather than a chance position; clearer cellular and subcellular detail; preservation of hydrated gels like mucilage and swollen cell walls; the possibility of making serial parallel sections through the same piece of tissue; opportunities for accurate morphometric analyses on the planed face; capacity to produce longitudinal sections; preservation of very delicate structures that are destroyed by fixation and drying. A major advantage of the Al-coated planed face is the increased accuracy of energy-dispersive X-ray (EDX) microanalyses on a smooth rather than a rough surface. Tests are included which show that neither the light etching employed, nor successive planing, interferes with the analyses of elements in the frozen face. © 1994 Wiley-Liss, Inc. IS - 1 JF - Microscopy Research and Technique AD - Cryo-Analytical SEM Facility, Carleton University, Ottawa, Canada K1S 5B6; Biology Department, Carleton University, Ottawa, Canada K1S 5B6; Department of Biological Sciences, Lancaster University, Lancaster LA1 4YQ, England EP - 74 TI - Planing frozen hydrated plant specimens for SEM observation and EDX microanalysis VL - 28 SP - 67 KW - cryo KW - techniques AU - Huang, Cheng AU - Canny, Martin AU - Oates, Kenneth AU - Mccully, Margaret PY - 1994/// UR - http://dx.doi.org/10.1002/jemt.1070280108 ER -