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Characterization of agarose as an encapsulation medium for particulate specimens for transmission electron microscopy
Author(s) -
Wood Jacqueline I.,
Klomparens Karen L.
Publication year - 1993
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.1070250402
Subject(s) - agarose , uranyl acetate , gelatin , osmium tetroxide , agar , staining , transmission electron microscopy , chemistry , materials science , chromatography , chemical engineering , electron microscope , nanotechnology , biochemistry , bacteria , biology , genetics , physics , engineering , optics
Agarose, agar, and gelatin were initially compared as encapsulation media for 3 structurally diverse particulate specimens: bacteria, yeast, and mitochondria. Agarose proved superior to both gelatin and agar for ease of handling and overall image quality (minimum background). All sample types exhibited high quality fixation and structural detail with no heat damage from the agarose medium. Based on this finding, we further characterized agarose encapsulation as affected by post‐fixation, en bloc staining and resin type. Osmium tetroxide post‐fixation, followed by en bloc uranyl acetate staining, could be performed without an increase in the electron density of the encapsulation medium. Agarose proved successful as an encapsulation medium regardless of the resin type or preparation protocol, thus providing flexibility in experimental design and excellent results over a range of variables. © 1993 Wiley‐Liss, Inc.