The authors studied the influence of fixations on the normal hydration of the rabbit cornea and the total amount of acid mucopolysaccharides (AMPS) in the stroma. The following fixatives were used: formol-calcium chloride at 19° C for 24 hours, formolcetylpyridinium chloride (CPC) at 19 and 28° C for 48 hours, Lillie's fixation at 19° C for 24 hours and Carnoy's fluid at 19° C for 30 minutes. The sections of the corneae were stained with Alcian blue, colloidal Fe3+ in the modification according to Rinehart and Abu'l Haj and with toluidine and methylene blue. The amount of AMPS was determined with the method of Rondle and Morgan and the total hydration of the stroma by weighing the corneae before and after using different fixative fluids and by calculation of obtained values on dry weight.
The best results were obtained by using formol-CPC at 28° C. At the ordinary room temperature (±19° C) it was the poorest fixation, however, as the corneae in this solution became hydrated. Formol-calcium chloride was the second in the row and it was much better than Lillie's and Carnoy's fluid.
The amount of AMPS in the stroma was not essentially changed by the effect of fixatives. Within 24–48 hours formol-CPC at 28° C retained the normal content and formol-calcium chloride caused the 11% decrease of AMPS maximally. The loss of AMPS after other fixatives was minimal.
The intensity of staining with cationic dyes in paraffin sections was different after individual fixatives and after the kind of their application and was dependent chiefly on the state of hydration of the corneal stroma: It is impossible to interpret the results of staining reactions in terms of the quantity of AMPS as it was hitherto done.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Anseth, A.: Studies on corneal polysaccharides. III. Topographic and comparative biochemistry. Exp. Eye Res. 1, 106–115 (1961).
Ashton, N.: The effect of swelling on the staining reactions of the cornea. In: The transparency of the cornea. A symposium organized by “The council for international organizations of medical sciences”. Edit. by Sir S. Duke-Elder and E. S. Perkins, p. 17–21. Oxford: Blackwell Sci. Publ. 1960.
Balazs, E. A.: The structure of the eye. Edit. by G. K. Smelser, p. 419. New York: Academic Press 1960.
Brewer, D. B.: Myxoedema: an autopsy report with histochemical observations on the nature of the mucoid infiltrations. J. Path. Bact. 63, 503–512 (1951).
Čejková, J., and I. Brettschneider: Hydration of rabbit cornea in vitro. Histochemical and biochemical study. (In preparation.)
—, u. J. Obenberger: Nachweis saurer Mucopolysaccharide in der Kaninchen-Hornhaut, fixiert nach verschiedenen Verfahren. Čs. Oftal. 24, 1–7 (1968).
Clemens, H.-J.: Über die Kittsubstanzen in der Propria corneae des Rindes. Acta histochem. (Jena) 1, 287–302 (1955).
Curran, R. C., and J. S. Kennedy: The distribution of the sulphated mucopolysaccharides in the mouse. J. Path. Bact. 70, 449–457 (1955).
Dempsey, E. W., H. Bunting, M. Singer, and G. B. Wislocki: The dyebinding capacity and other chemo-histological properties of mammalian mucopolysaccharides. Anat. Rec. 98, 417–428 (1947).
Dohlman, C. H.: Incorporation of radioactive sulfate into the rabbit eye. Acta ophthal. (Kbh.) 35, 115–124 (1957).
Ehlers, N.: Studies on the hydration of the cornea with special reference to the acid hydration. Acta ophthal. (Kbh.) 44, 924–931 (1966).
Francois, J. F., et M. R. Rabaey: Recherches histo-chimiques sur le mucoide de la cornée. Ann. Oculist. (Paris) 186, 901–924 (1953).
— Nouvelles recherches physico- et histo-chimiques sur le mucoide de la cornée. Acta ophthal. (Kbh.) 34, 45–62 (1956).
Graumann, W., u. J. Rohen: Chemohistologische Befunde am menschlichen Auge. (Cornea, Sclera, Uvea.) Z. mikr. anat. Forsch. 64, 652–671 (1958).
Hale, C. W.: Histochemical demonstration of acid polysaccharides in animal tissues. Nature (Lond.) 157, 802 (1946).
Harris, J. E., and L. T. Nordquist: The hydration of the cornea. I. The transport of water from the cornea. Amer. J. Ophthal. 40, pt. II, 100–110 (1955).
Kern, H. L., and W. M. Grant: Interaction of acidic and basic dyes and beef corneal stroma. J. Histochem. Cytochem. 9, 380–385 (1961).
Larsen, G.: An autoradiographic study on the uptake of S35-labelled sodium sulfate. In the eyes of normal, scorbutic and hormonal treated guinea pig. Amer. J. Ophthal. 47, part II, 519–529 (1959).
Lillie, R. D.: Histopathologic technic and practical histochemistry. Philadelphia: Blakiston 1954.
Lojda, Z.: Fixation in histochemistry. Folia morph. (Prague) 13, 65–83 (1965a).
- Histochemistry of the vascular wall. In: Int. Symp. Morphology Histochemistry Vascular Wall, Fribourgh 1965, part II, 364–398 (1965b).
McManus, J. F. A.: Histological and histochemical uses of periodic acid. Stain Technol. 23, 99–108 (1948).
—, and R. W. Mowry: Effects of fixation on carbohydrate histochemistry. J. Histochem. Cytochem. 6, 309–316 (1958).
Nicholas, J. P.: Mucopolysaccharide staining in experimental corneal edema. Arch. Ophthal. 76, 238–243 (1966).
Pearse, A. G. E.: Histochemistry, theoretical and applied. Boston: Little, Brown & Co. 1961.
Rinehart, J. F., and S. K. Abu'l-Haj: An improved method for histologic demonstration of acid mucopolysaccharides in tissues. Arch. Path. 52, 189–194 (1951).
Rondle, C. J. M., and W. J. M. Morgan: The determination of glucosamine and galactosamine. Biochem. J. 61, 586–589 (1955).
Sochor, F. M.: Effect of various fixatives on dissolution and staining characteristics of acid mucopolysaccharides in human aorta. Amer. J. clin. Path. 44, 636–638 (1965).
Spicer, S. S.: Histochemical properties of mucopolysaccharides and basic protein in mast cells. Ann. N.Y. Acad. Sci. 103, 322–333 (1963).
Szirmai, J. A.: Quantitative approaches in the histochemistry of mucopolysaccharides. J. Histochem. Cytochem. 11, 24–34 (1963).
Williams, G., and D. S. Jackson: Two organic fixatives for acid mucopolysaccharides. Stain Technol. 31, 189–191 (1956).
Zugibe, F. T.: Mucopolysaccharides of the arterial wall. J. Histochem. Cytochem. 11, 35–39 (1963).
About this article
Cite this article
Čejková, J., Brettschneider, I. On the influence of fixations on the normal hydration of rabbit cornea and the total amount of acid mucopolysaccharides in the stroma. Histochemie 17, 108–120 (1969). https://doi.org/10.1007/BF00277776
- Methylene Blue
- Paraffin Section
- Normal Content