Enhanced ultrastructural preservation of cartilage proteoglycans in the extended state

We investigated the preservation of proteoglycan (PG) structure in rat epiphyseal cartilage using N-N-dimethylformamide (DMF) dehydration before embedding. After aldehyde fixation, specimens with and without routine osmium post-fixation were dehydrated in graded DMF and embedded in either Spurr'...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1990-06, Vol.38 (6), p.901-906
Main Authors: Kagami, A, Takagi, M, Hirama, M, Sagami, Y, Shimada, T
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell physiology
Dimethylformamide
Fundamental and applied biological sciences. Psychology
Growth Plate - ultrastructure
Male
Molecular and cellular biology
Preservation, Biological - methods
Protein Conformation
Proteoglycans - ultrastructure
Rats
Rats, Inbred Strains
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Summary:We investigated the preservation of proteoglycan (PG) structure in rat epiphyseal cartilage using N-N-dimethylformamide (DMF) dehydration before embedding. After aldehyde fixation, specimens with and without routine osmium post-fixation were dehydrated in graded DMF and embedded in either Spurr's resin or Lowicryl K4M resin. Standard ethanol dehydration with Spurr or Lowicryl embedding techniques resulted in the formation of condensed PGs, called matrix granules. DMF dehydration before embedding greatly improved the preservation of PG structure and resulted in an extended appearance of PGs closely resembling the fine filamentous network of cartilage tissues processed by rapid freezing and freeze-substitution. However, en bloc staining of aldehyde-fixed specimens with cationic reagents before or during DMF dehydration induced the condensation of PGs and resulted in the formation of matrix granules. These observations demonstrate that DMF, a mild dehydration agent, dramatically improves PG preservation without a harmful effect on aldehyde-fixed PG structure and can be utilized regardless of routine post-fixation.
ISSN:0022-1554
1551-5044
DOI:10.1177/38.6.2335743