Summary
Two new histochemical procedures for detecting sulphated and non-sulphated sialomucin in colonic mucosa were assessed: the saponification—Alcian Blue pH 1—periodic acid—phenylhydrazine—Schiff method (KOH—AB pH 1—PAPS) and the mild periodic acid modification of this (KOH—AB pH 1—mPAS). Using normal colonic mucosa obtained from 11 non-cancer patients, the mPAS and PAPS techniques were tested for specificity and reproducibility for staining sialic acid, either alone or in combination with Alcian Blue. A spectrophotometric method was devised to quantify the uptake of both Schiff and Alcian Blue stain by sections. At low temperature and pH5.5, the mPAS procedure had improved specificity over the PAPS procedure, and after saponification it could be used to stainO-acetyl-substituted sialic acid. When used in combination with Alcian Blue at pH 1, however, underestimation of the sialic acid content occurred owing to interference between Alcian Blue and Schiff dyes. Interference was even greater with KOH—AB pH1—PAPS procedure for both sialic acid and sulphate components. We conclude that caution must be exercised in interpretation of the staining results obtained with these new combination methods and that more accurate information on the sialic acid and sulphate content of colonic mucin is obtained by staining serial sections with the mPAS technique and Alcian Blue pH 1 alone.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
BANCROFT, J. D. & STEVENS, A. (1982) Preparation of Schiff's reagent. InTheory and Practice of Histological Techniques (edited by BANCROFT, J. D. & STEVENS, A.), pp. 189–90. London: Churchill Livingstone.
BUK, S. J. A. & FILIPE, M. I. (1986) Histochemical staining of sulphated and non-sulphated sialomucin in intestinal epithelium: practical difficulties associated with the KOH.AB.1.0.PAPS method.Histochem. J. 18, 576–8.
CORFIELD, A. P. & ROE, R. (1988) Spectrophotometric analysis of histochemical staining. InSpectroscopic and Structural Studies of Materials and Systems of Fundamental Importance to Biology and Medicine. IV. Biomolecular Interactions. Sigma Press (in press).
FILIPE, M. I. & BRANFOOT, A. C. (1974) Abnormal patterns of mucous secretion in apparently normal mucosa of large intestine with carcinoma.Cancer 34, 282–90.
MCFADDEN, D. E., OWEN, D. A., REID, P. E. & JONES, E. A. (1985) The histochemical assessment of sulphated and non-sulphated sialomucin in intestinal epithelium.Histopathology 9, 1129–37.
REID, P. E., CULLING, C. F. A., DUNN, W. L., RAMEY, C. W. & CLAY, M. G. (1984a) Chemical and histochemical studies of normal and diseased human gastrointestinal tract: I. A comparison between histologically normal colon, colonic tumours, ulcerative colitis and diverticular disease of the colon.Histochem. J. 16, 235–51.
REID, P. E., CULLING, C. F. A., DUNN, W. L., RAMEY, C. W., MAGIL, A. B. & CLAY, M. G. (1980) Differences between theO-acetylated sialic acids of the epithelial mucins of human colonic tumours and normal controls. A correlative chemical and histochemical study.J. Histochem. Cytochem. 28, 217–22.
REID, P. E., DUNN, W. L., RAMEY, C. W., CORET, E., TRUEMAN, L. & CLAY, M. G. (1984b) Histochemical studies of the mechanism of the periodic acid—phenyl-hydrazine—Schiff (PAPS) procedure.Histochem. J. 16, 641–9.
REID, P. E., OWEN, D. A., RAMEY, C. W., DUNN, W. L., CLAY, M. G. & JONES, E. A. (1985) Histochemical procedures for the simultaneous visualisation of sialic acid, its side chainO-acyl variants andO-sulphate esters.Histochem. J. 17, 113–17.
ROBERTS, G. P. (1977) Histochemical detection of sialic acid residues using periodate oxidation.Histochem. J. 9, 97–102.
ROGERS, C. M., CORFIELD, A. P., REUTER, G., FILIPE, M. I., COOKE, K. B. & SCHAEUR, R. (1979) A micromethod for the preparation and analysis of sialic acids from normal and malignant human large bowel mucosa. InGlycoconjugates (edited by SCHAUER, R., BOER, P., BUDDECKE, E., KRAMER, M. F., VLIEGENTHART, J. F. G. & WIEGANDT, H.). pp. 652–3, Stuttgart: Georg Thieme.
SHUKLA, A. K. & SCHAUER, R. (1982) Study of de-O-acetylation ofN-acetyl-9-O-acetyl neuraminic acid by a fluorimetric method.Fresenius Z. Anal. Chem. 311, 377.
SPICER, S. S. (1961) The use of various cationic reagents in histochemical differentiation of mucopolysaccharides.Amer. J. Clin. Pathol. 36, 393–409.
SPICER, S. S. (1965) Diamine methods for differentiating mucosubstances histochemically.J. Histochem. Cytochem. 13, 211–34.
SUGIHARA, K. & JASS, J. R. (1986) Colorectal goblet cell sialomucin heterogeneity: its relation to malignant disease.J. Clin. Pathol. 39, 1088–95.
VARKI, A. & DIAZ, S. (1984) The release and purification of sialic acids from glycoconjugates: methods to minimize the loss and migration ofO-acetyl groups.Anal. Biochem. 137, 236–47.
VEH, R. W., CORFIELD, A. P., SCHAUER, R. & ANDRES, K. H. (1979) The bovine submandibular gland. II. Histochemical and biochemical results. In Glycoconjugates (edited by SCHAUER, R., BOER, P., BUDDECKE, E., KRAMER, M. F., VLIEGENTHART, J. F. G. & WIEGANDT, H.), pp. 193–4. Stuttgart: Georg Thieme.
VEH, R. W., MEESEN, D. KUNTZ, H. D. & MAY, B. (1982) Histochemical demonstration of side-chain substituted sialic acids. InColonic Carcinogenesis (Falk Symposium 31) (edited by MALT, R. A. & WILLIAMSON, R. C. N.), pp. 355–65. Lancaster: MTP press.
WEBER, P., HARRISON, F. W. & HOF, L. (1975) Histochemical application of dansylhydrazine as a fluorescent labelling reagent for sialic acid in cellular glycoconjugates.Histochemistry 45, 271–7.
WILLIAMS, G. T. (1985) Transitional mucosa of the large intestine: commentary.Histopathology 9, 1243–5.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Roe, R., Corfield, A.P. & Williamson, R.C.N. Sialic acid in colonic mucin: An evaluation of modified PAS reactions in single and combination histochemical procedures. Histochem J 21, 216–222 (1989). https://doi.org/10.1007/BF01747523
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01747523