Synopsis
In this paper, it is assumed that tissue fixation is a process in which the proteins become less soluble and catabolic reactions stop. With this definition in mind, 2.5 and 5% glutaraldehyde and 4% formaldehyde ino.i m potassium phosphate buffer, pH 7.4, were compared with a new fixative, bicarbonate-formaldehyde. The following results were obtained. (1) With 2.5 and 5% glutaraldehyde, the solubility of tissue proteins were not decreased uniformly, and tissue glycogen was poorly preserved. (2) 4% formaldehyde in potassium phosphate buffer gave relatively good results. (3) Bicarbonate-formaldehyde decreased the solubility of tissue proteins reliably and preserved tissue glycogen perfectly. Histologically, it yielded excellent results.
Since glutaraldehyde alters the properties of proteins substantially (Hopwood, 1972; Habeeb & Hiramoto, 1968), and since the natural appearance of tissues depends on native tissue proteins, formaldehyde-containing fixatives, in particular bicarbonate-formaldehyde, are preferable to glutaraldehyde-containing fixatives for all tissue preparative techniques. However, it is important that the fixation time in formaldehyde is kept short.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abramezuk, J. (1972). Effect of formalin fixation on the dry mass of isolated rat liver nuclei.Histochemie 29, 207–12.
Baker, J. R. &Mccrae, J. M. (1966). The fine structure resulting from fixation by formaldehyde: The effects of concentration, duration, and temperature.Jl. R. microsc. Soc. 85, 391–4.
Bone, Q. &Denton, E. J. (1971). The osmotic effects of electron microscope fixatives.J. Cell Biol. 49, 571–81.
Bowes, J. A. &Cater, C. W. (1968). The interaction of aldehydes with collagen.Biochim. biophys. Acta 168, 341–52.
Cantarow, A. &Schepartz, B. (eds.) (1967).Biochemistry, p. 308. Philadelphia, London: Saunders.
Chambers, R. W., Bowling, M. C. &Grimley, P. M. (1968). Glutaraldehyde fixation in routine histopathology.Arch. Path. 85, 18–30.
Dawson, I. M. P. (1972), Fixation: What should the pathologist do?.Histochem. J. 4, 382–3.
Dimodica, G. &Marzona, M. (1971). Crosslinking of wool keratin by bifunctional aldehydes.Textile Res. J. 41, 701–5.
Ellar, D. J., Munoz, E. &Salton, M. R. (1971). The effect of low concentrations of glutaraldehyde onMicrococcus lysodeikticus membranes: changes in the release of membrane associated enzymes and membrane structure.Biochim. biophys. Acta 225, 140–50.
Ericsson, J. L. E. &Biberfeld, P. (1967). Studies on aldehyde fixation. Fixation rates and their relation to fine structure and some histochemical reactions in liver.Lab. Invest. 17, 281–98.
Flitney, E. W. (1966). The time course of fixation of albumin by formaldehyde, glutaraldehyde, acrolein and other higher aldehydes.Jl. R. microsc. Soc. 85, 353–64.
Crillo, T. A. I., Ogunnaire, P. O. &Faoye, S. (1971). Effects of histological and electron microscopical fixatives on the insulin content of the rat pancreas.J. Endocr. 51, 645–9.
Habeeb, A. F. S. A. &Hiramoto, R. (1968). Reaction of protein with glytaraldehyde.Arch. Biochem. Biophys. 126, 16–22.
Höber, R. (1945). The surface of the protoplast, its properties and its architecture. In:Physical Chemistry of Cells and Tissues (eds. R. Höber, D. I. Hitchcock, J. R. Bateman, D. R. Goddard & W. O. Fenn), p. 246. Philadelphia, Toronto: Blakiston.
Hopwood, D. (1967a). Some aspects of fixation with glutaraldehyde. A biochemical and histochemical comparison of the effects of formaldehyde and glutaraldehyde fixation on various enzymes and glycogen with a note on penetration of glutaraldehyde into liver.J. Anat. 101, 83–92.
Hopwood, D. (1967b). The effect of formaldehyde fixation and dehydration on ox adrenal medulla with respect to the chromaffin reaction and post-chroming.Histochemie 10, 98–106.
Hopwood, D. (1969). A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and α-hydroxydipaldehyde with bovine serum albumin and casein.Histochemie 17, 151–61.
Hopwood, D. (1970). The reactions between formaldehyde, glutaraldehyde and osmium tetroxide and their fixation effects on bovine serum albumin and on tissue blocks.Histochemie 26, 56–64.
Hopwood, D., Allen, C. R. &Mccabe (1970). The reactions between glutaraldehyde and various proteins. An investigation of their kinetics.Histochem. J. 2, 137–50.
Hopwood, D. (1971). Use of isoelectric focusing to determine the isoelectric point of bovine serum albumin after treatment with various common fixatives.Histochem. J. 3, 201–5.
Hopwood, D. (1972). Theoretical and practical aspects of glutaraldehyde fixation.Histochem. J. 4, 267–303.
Huggett, A. St. G. &Nixon, D. A. (1957). Use of glucose oxidase, peroxidase and O-dianisidine in determination of blood and urinary glucose.Lancet ii, 368.
Jansen, E. F., Tomimatsu, Y. &Olsen, A. C. (1971). Crosslinking of α-chymotrypsin and other proteins by reaction with glutaraldehyde.Arch. Biochem. Biophys. 144, 394–400.
Joly, M. (1965). A physico-chemical approach to the denaturation of proteins.Molecular Biology, an International Series of Monographs and Textbooks (eds. B. Horecker, N. O. Kaplan & H. A. Scheraga), Vol. 6, p. 162. London, New York: Academic Press.
Jozsa, L. &Szederkenyi, G. (1967). Loss of mucopolysaccharides from tissue during fixation.Acta Histochem. 26, 255–60.
Layne, E. (1957). In:Methods in Enzymology (eds. S. P. Colowick & N. O. Kaplan), Vol. 3, pp. 450–4. New York: Academic Press.
Lenard, J. &Singer, S. J. (1968). Alterations of the conformation of proteins in red blood cell membranes and in solution by fixatives used in electron microscopy.J. Cell Biol. 37, 117–22.
Ottesen, M. &Svensson, B. (1971). Modification of papain by treatment with glutaraldehyde under reducing and non-reducing conditions.C. R. Lab. Carlsberg 38, 171–85.
Pease, D. C. (1964).Histological Techniques for Electron Microscopy, New York, London: Academic Press.
Roux, S. J. &Hillman, W. S. (1969). The effect of glutaraldehyde and two monoaldehydes on phytochrome.Arch. Biochem. Biophys. 131, 423–9.
Somogyi, M. (1957). In:Methods in Enzymology (eds. S. P. Colowick & N. O. Kaplan), Vol. 3, p. 3. New York: Academic Press.
Stecher, P. G., Finkel, M. J., Siegmund, O. H. &Szafranski, B. M. (eds.) (1960).The Merck Index of Chemicals and Drugs, p. 946. Rahway, N.J., U.S.A.: Merck.
Thompson, S. W. (1966).Selected Histochemical and Histopathological Methods. Springfield, Illinois U.S.A.: Charles C. Thomas.
Wold, F. (1961). The reaction of bovine serum albumin with the fibunctional reagentp′,p-difluoro-m,m′-dinitro-diphenylsulfone.J. Biol. Chem. 236, 106–11.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Artvinli, S. Biochemical aspects of aldehyde fixation and a new formaldehyde fixative. Histochem J 7, 435–450 (1975). https://doi.org/10.1007/BF01003881
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01003881