Ultrastructure of Kurloff body proteoglycans after high pressure freezing, cryosubstitution and postembedding staining with cuprolinic blue

Very high pressure freezing and cryosubstitutlon of Kurloff cells preserves the ultrastructural morphology of Kurloff bodies, particularly the myelin figures, as shown by embedding in epoxy resin and conventional postembedding staining. It also preserves the Kurloff body proteoglycans as more expand...

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Bibliographic Details
Published in:	Glycobiology (Oxford) Vol. 6; no. 8; pp. 817 - 822
Main Authors:	Landemore, Gérard, Quillec, Michèle, Izara, Jacques
Format:	Journal Article
Language:	English
Published:	England Oxford University Press 01-12-1996
Subjects:	Animals Coloring Agents cuprolinic blue Freezing Guinea Pigs high pressure freezing Indoles Kurloff cell Microscopy, Electron Organometallic Compounds proteoglycans Proteoglycans - ultrastructure Spleen - metabolism Spleen - ultrastructure Staining and Labeling ultrastructure
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Summary:	Very high pressure freezing and cryosubstitutlon of Kurloff cells preserves the ultrastructural morphology of Kurloff bodies, particularly the myelin figures, as shown by embedding in epoxy resin and conventional postembedding staining. It also preserves the Kurloff body proteoglycans as more expanded spindle-like shapes than does fixation with formaldehyde at atmospheric pressure. But., proteoglycans were not discernible in the Kurloff body matrix on either unstained or conventionally stained thin sections. The Kurloff body skeleton of proteoglycans in their native expanded shape was stained with the electron-dense cationic ministain cuprolinic blue, using thin sections embedded in LR white. The mean equatorial diameter of the spindles was 20–30 nm, while the collapsed filaments produced by aldehyde fixation were about 10–15 nm wide. The spindles were often about 200–300 nm long but could be much longer, depending on the plane of the section. Thus, high pressure freezing, freeze substitution, embedding in LR white, and staining with cationic dyes such as phthalocyanins seems to be a convenient way of visualizing intracellular proteoglycans that are well preserved and in very much like their native expanded state.
Bibliography:	ArticleID:6.8.817 istex:879728D9F3B412EEF7171D33FC39372D42C69643 ark:/67375/HXZ-6CX72MQF-6 2To whom correspondence should be addressed ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0959-6658 1460-2423
DOI:	10.1093/glycob/6.8.817