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Application of ion etching to immunoscanning electron microscopy
Author(s) -
Yahiro Junko,
Nagato Toshikazu
Publication year - 2005
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.20202
Subject(s) - immunogold labelling , scanning electron microscope , electron microscope , materials science , scanning confocal electron microscopy , etching (microfabrication) , chemistry , secondary electrons , analytical chemistry (journal) , microscopy , electron , optics , nanotechnology , chromatography , composite material , physics , layer (electronics) , quantum mechanics
A method for achieving both the light and electron microscopic observations of the same immunolabeled semithin section is described. Mild ion etching (IE) was performed on the semithin LR white resin sections of rat pancreas to evaluate conditions for scanning electron microscopic secondary electron image observations. Before immunocytochemical staining, very mild, rapid etching was conducted as follows: ionization voltage 300 V, operating vacuum 35 Pa, and etching time 1 min, employing an ion coater above sections on glass slides. The sections were immunohistochemically stained with anti‐insulin and immunogold in association with silver enhancement techniques for light microscopic observation, in which B cells in pancreatic islets were positively stained brown. Subsequently, essential mild IE was performed over the stained section as follows: 350 V, 38 Pa, 29 min. The samples were coated with platinum for scanning electron microscopic secondary electron images, in which the cores of secretory granules of the B cells were positively labeled with gold–silver particles. The present method is suitable for detection of substances involving immunogold labeling. It enables us to obtain high‐resolution images at low magnification that can be correlated with light microscopic observations. Middle to high magnifications are applicable for detailed observations with secondary electron imaging scanning electron microscopy. Microsc. Res. Tech. 67:240–247, 2005. © 2005 Wiley‐Liss, Inc.