Immune electron microscopy using glutaric-fixed epoxy resin-embedded block (Yano/Kajima method).
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Accession number;05A0266881
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| Title;Immune electron microscopy using glutaric-fixed epoxy resin-embedded block (Yano/Kajima method). |
| Author;
KASHIMA KENJI
(Oitadai I Byoin Byoribu)
YANO SHINJI
(Oita Univ.)
DAA TSUTOMU
(Oitadai I Byoin Byoribu)
YOKOYAMA SHIGEO
(Oita Univ.)
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Journal Title;Modern Medical Laboratory
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Journal Code:Z0084B
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ISSN:0301-2611
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VOL.33;NO.2;PAGE.111-114(2005)
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| Figure&Table&Reference;FIG.3, REF.3 |
| Pub. Country;Japan |
| Language;Japanese |
| Abstract;When a sample for immune electron microscopy is to be prepared, it is a problem that only the epoxy embedding block for usual electron microscopy is generally produced. Moreover, in this block, the glutaric fixation/OsO4 postfixation with a strong effect on the antigenicity has been applied. Therefore, antigen activation by heating was tried for an ultrathin slice produced from this block, and a result which can be practiced in the observation by immune electron microscopy was obtained. It is explained in the order of 1) necessary instrument/reagent, 2) staining method, 3) staining agent, and 4) application cases (case in which target cells are few, case with only paraffin block or paraffin slice). In the technique, to begin with, the ultrathin slice is picked up on a nickel grid knife. The slice, together with the grid, is dipped in "target retrieval solution, high pH", an antigen activating solution, and after heating by electronic range for 15 minutes, is exposed to the room temperature. It is washed with Tris-HCl buffer solution added with 0.1% BSA and 0.1% tween 20, and after immunostaining, uranil staining and lead staining are performed, it is observed by an electron microscope. By this method, it is possible to identify secretory granules, various peptide hormones and human papilloma virus. |
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