Summary
Neutral buffered 4% formaldehyde fixation for 48h preserved well the proteoglycan content of bovine articular cartilage. Neither subsequent demineralization in 10% EDTA, nor light microscopic tissue processing, reduced the hexosamine or uronic acid content of the tissue. Fixation in alcoholic solutions increased Safranin O binding as well as periodic acid Schiff reaction of the cartilage matrix as measured by microspectrophotometry. It is suggested that the enhanced staining of cartilage was due to better preservation of the glycoprotein oligosaccharides. Quaternary ammonium salts in the fixative suppressed the staining of cartilage matrix with Safranin O.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Allison RT (1973) The effects of fixation on subsequent demonstration of mucopolysaccharides. Med Lab Technol 30:27–31
Blumenkranz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54:484–489
Boas NF (1953) Method for the determination of hexosamines in the tissues. J Biol Chem 204:553–563
Charman J, Reid L (1972) The effect of decalcifying fluids on the staining of epithelial mucins by Alcian Blue. Stain Technol 47:173–178
Conklin JL (1963) Staining reactions of mucopolysaccharides after formalin-containing fixatives. Stain Technol 38:56–59
Cook HC (1977) Carbohydrates. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques. Churchill Livingstone, Edinburgh London New York, pp 141–167
Eggert FM, Germain JP (1979) Rapid demineralization in acidic buffers. Histochemistry 59:215–224
Eggert FM, Linder JE, Jubb RW (1981) Staining of demineralized cartilage. I. Alcoholic versus aqueous demineralization at neutral and acidic pH. Histochemistry 73:385–390
Engfeldt B, Hjertquist S-O (1967) The effect of various fixatives on the preservation of acid glycosaminoglycans in tissues. Acta Pathol Microbiol Scand 171:219–232
Fejerskov O (1971) The effect of different demineralizing agents on oral mucous membrane. Scand J Dent Res 79:172–182
Hopwood D (1977) Fixation and fixatives. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques. Churchill Livingstone, Edinburgh London New York, pp 16–28
Hopwood D, Mellors AE (1973) An investigation of the chlorotriazine derivatives as fixatives for tissues. Histochem J 5:391–403
Horobin RW (1982) Histochemistry. An explanatory outline of histochemistry and biophysical staining. Gustav Fisher, Stuttgart New York; Butterworths, London Boston Syndney Wellington Durban Toronto
Józsa L, Szederkényi G (1967) Über verluste der Gewebsmukopolysaccharide während der Fixierung. Acta Histochem 26:255–260
Jubb RW, Eggert FM (1981) Staining of demineralized cartilage II. Quantitation of articular cartilage proteoglycan after fixation and rapid demineralization. Histochemistry 73:391–396
Kiviranta I, Tammi M, Lappalainen R, Kuusela T, Helminen HJ (1980) The rate of calcium extraction during EDTA decalcification from thin bone slices as assessed with atomic absorption spectrophotometry. Histochemistry 68:119–127
Knese K-H (1977) The fixation of intracellular carbohydrates in the chondroxytes. Acta Histochem 59:139–154
Korhonen LK, Lindholm K (1969) Effect of histological fixation on carbohydrate-rich tissue compounds and their periodate consumption. Histochemie 18:87–96
Pearse AGE (1980) Histochemistry. Theoritical and applied. Vol. 1. Preparative and optical technology, 4th ed. Churchill Livingstone, Edinburgh London New York, p 140
Schajowicz F, Cabrini RL (1955) The effect of acids (decalcifying solutions) and enzymes on the histochemical behavior of bone and cartilage. J Histochem Cytochem 3:122–129
Scott JF, Dorling J (1969) Periodate oxidation of acid polysaccharides III. A PAS method for chondroitin sulphates and other glycosaminoglycuronans. Histochemie 19:295–301
Scott JE (1955) The solubility of cetylpyridinium complexes of biological polyanions in solution of salts. Biochim Biophys Acta 18:428–429
Sochor FM (1965) Effect of various fixatives on dissolution and staining characteristics of acid mucopolysaccharides in human aorta. Am J Clin Pathol 44:636–638
Szirmai JA (1963) Quantitative approaches in the histochemistry of mucopolysaccharides. J Histochem Cytochem 11:24–34
Torrello LA, Yates AJ, Thompson DK (1980) Critical study of the alditol acetate method for quantitating small quantities of hexoses and hexosamines in gangliosides. J Chromatogr 202:195–209
Tyrkkö J, Häkkinen I, Rimpelä U (1968) On the histochemical demonstration of sulphomucins in human and canine gastric epithelium. Br J Exp Pathol 49:371–374
Williams G, Jackson DS (1956) Two organic fixatives for acid mucopolysaccharides. Stain Technol 31:189–191
Wolman M (1955) Problems of fixation in cytology, histology, and histochemistry. Int Rev Cytol 4:79–102
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kiviranta, J., Tammi, M., Jurvelin, J. et al. Fixation, decalcification, and tissue processing effects on articular cartilage proteoglycans. Histochemistry 80, 569–573 (1984). https://doi.org/10.1007/BF00553719
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00553719