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Theoretical and practical aspects of glutaraldehyde fixation

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Synopsis

This review first considers the many structures put forward for glutaraldehyde, and the purification of the commercial material for chemical, histological and histochemical studies. Some practical and theoretical problems of tissue fixation with glutaraldehyde, including artefacts, are then discussed. The chemical reactions with amino acids and proteins are considered next together with the physical changes in the proteins during the reactions. The known reactions of glutaraldehyde with nucleic acids, lipids and mucosubstances are explored briefly.

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References

  • Alexa, G., Chisalita, D. &Chirita, G. (1971). Reaction of dialdehyde with functional groups in collagen.Rev. Tech. Ind. Cuir. 63, 5–6.

    Google Scholar 

  • Amsterdam, A. &Schramm, M. (1966). Rapid release of zymogen granule protein by osmium tetroxide and its retention by glutaraldehyde.J. Cell Biol. 29, 199–207.

    Google Scholar 

  • Anderson, P. J. (1967). Purification and quantitation of glutaraldehyde and its effects on several enzyme activities in skeletal muscle.J. Histochem. Cytochem. 15, 652–61.

    Google Scholar 

  • Arborgh, B., Ericsson, J. L. E. &Helminen, H. (1971). Inhibition of renal acid phosphatase and arylsulphatase activity by glutaraldehyde fixation.J. Histochem. Cytochem. 19, 449–51.

    Google Scholar 

  • Aso, C. &Aito, Y. (1962). Studies on the polymerization of bifunctional monomers. II polymerization of glutaraldehyde.Makromol. Chem. 58, 195–203.

    Google Scholar 

  • Axén, R., Myrin, P. Å. &Jansson, J. C. (1970). Chemical fixation of chymotrypsin to water insolubilized cross linked Dextran (Sephadex) and solubilization of the enzyme derivatives by means of a dextranase.Biopolymers 9, 401–13.

    Google Scholar 

  • Bahr, G. F. (1954). Osmium tetroxide and ruthenium tetroxide and their reactions with biologically important substances.Exp. Cell Res. 7, 457–89.

    Google Scholar 

  • Baker, J. R. (1960).Principles of Biological Microtechnique. London: Methuen.

    Google Scholar 

  • Beauvillain, J. C. (1970). Influence de divers fixateurs sur la structure fine de L'éminence médiane de Souris.C. R. Soc. Biol. 164, 1032–34.

    Google Scholar 

  • Berns, K. I. &Thomas, C. A. (1961). A study of single polynucleotide chains derived from T2 and T4 bacteriophage.J. Mol. Biol. 3, 289–300.

    Google Scholar 

  • Bishop, W. H. &Richard, F. M. (1968). Isoelectric point of a protein in the cross linked state:-lactoglobulin.J. Mol. Biol. 33, 415–21.

    Google Scholar 

  • Bloem, E. (1968). Thin layer chromatography of aldehydes in leather.J. Soc. leath. Trades Chem. 52, 204–7.

    Google Scholar 

  • Blumenfeld, O. O., Paz, M. A., Gallop, P. M. &Seifter, S. (1963). The nature, quantity and mode of attachment of hexoses in ichthyocol.J. biol. Chem. 238, 3835–9.

    Google Scholar 

  • Bodian, D. (1970). An electron microscopic characterization of synaptic vesicles by means of controlled aldehyde fixation.J. Cell Biol. 44, 115–24.

    Google Scholar 

  • Bohman, S. O. &Maunsbach, A. B. (1970). Effects on tissue fine structure of variations in colloid osmotic pressure of glutaraldehyde fixatives.J. Ultrastruct. Res. 30, 195–208.

    Google Scholar 

  • Bone, Q. &Denton, E. J. (1971). The osmotic effects of electron microscope fixatives.J. Cell Biol. 49, 571–81.

    Google Scholar 

  • Bone, Q. &Ryan, K. P. (1972). Osmolarity of osmium tetroxide and glutaraldehyde fixatives.Histochem. J. 4, 331–47.

    Google Scholar 

  • Bowes, J. A. &Cater, C. W. (1965). Cross linking of collagen.J. Appl. Chem. (London)15, 296–304.

    Google Scholar 

  • Bowes, J. A. &Cater, C. W. (1968). The interaction of aldehydes with collagen.Biochem biophys. Acta. 168, 341–52.

    Google Scholar 

  • Brooks, B. R. &Klamerth, O. L. (1968). Interaction of DNA with bifunctional aldehydes.Europ. J. Biochem. 5, 178–82.

    Google Scholar 

  • Carstensen, E. L., Aldridge, W. G., Child, S. Z., Sullivan, P. &Brown, H. H. (1971). Stability of cells fixed with glutaraldehyde and acrolein.J. Cell. Biol. 50, 529–32.

    Google Scholar 

  • Casey, E. J. (1962).Biophysics: Concepts and Mechanisms. New York: Reingold.

    Google Scholar 

  • Cejkova, J. &Brettschneider, I. (1970). Glutaraldehyde fixation of corneal acid mucopolysaccharides.Ophthalmic Res. 1, 149–55.

    Google Scholar 

  • Chambers, R. W., Bowling, M. C. &Grimley, P. M. (1968). Glutaraldehyde fixation in routine histopathology.Archs. Path. 85, 18–30.

    Google Scholar 

  • Chiang, J., Kowada, M., Ames, A., Wright, R. L. &Majno, G. (1968). Cerebral ischaemia III vascular changes.Am. J. Path. 52, 455–65.

    Google Scholar 

  • Chisalita, D., Chirita, G. &Alexa, G. (1971). Kinetics of dialdehyde combination with the reactive groups of collagen.Ind. Usoara 18, 269–81 (Roum).

    Google Scholar 

  • Clark, M. A. &Ackerman, G. A. (1971). Alteration of nuclear and nucleolar pyroantimonateosmium reactivity by glutaraldehyde fixation.J. Histochem. Cytochem. 19, 388–90.

    Google Scholar 

  • Cleland, W. W. (1967). In:Biological Chemistry (eds. H. R. Mahler & E. H. Cordes), p. 224. London: Harper & Row.

    Google Scholar 

  • Clift, F. P. &Cook, R. P. (1932). A method of determination of some biologically important aldehydes and ketones with special reference to pyruvic acid and methyl glyoxal.Biochem. J. 26, 1788–99.

    Google Scholar 

  • Coupland, R. E. &Hopwood, D. (1966). The mechanism of the differential staining reaction for adrenaline and noradrenaline storing granules in tissues fixed in glutaraldehyde.J. Anat. 100, 227–43.

    Google Scholar 

  • Curgy, J. J. (1968). Influence du mode de fixation sur la possibilité d'observer des structures myéliniques dans les hépatocytes d'embryons de poulet.J. Microscopie 7, 63–80.

    Google Scholar 

  • Daneel, S. &Weissenfels, N. (1965). Besseres Fixierungsverfahren zur Darstellung des Grundplasmas von Protozoen und Vertebraten zellen.Mikroskopie 20, 162–4.

    Google Scholar 

  • Deamer, D. W. &Crofts, A. (1967). Action of Triton X-100 on chloroplast membranes.J. Cell Biol. 33, 395–410.

    Google Scholar 

  • De Jong, D. W., Olson, A. C. &Jansen, E. F. (1967). Glutaraldehyde activation of nuclear acid phosphatase in cultured plant cells.Science 155, 1672–4.

    Google Scholar 

  • Descarries, L. &Droz, B. (1970). Intraneural distribution of exogenous norepinephine in the central nervous system of the rat.J. Cell Biol. 44, 385–99.

    Google Scholar 

  • di Modica, G. &Marzona, M. (1971). Cross linking of wool keratin by bifunctional aldehydes,Textile Res. J. 41, 701–5.

    Google Scholar 

  • Donnelly, E. H. &Goldstein, I. J. (1970). Glutaraldehyde insolubilized Concanavalin A: an adsorbent for the specific isolation of polysaccharides and glycoproteins.Biochem. J. 118, 679–80.

    Google Scholar 

  • 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.Biochem. biophys. Acta 225, 140–50.

    Google Scholar 

  • 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.

    Google Scholar 

  • Ernst, S. A. &Philpott, C. W. (1970). Preservation of Na-K-activated and Mg-activated ATP-ase activities of avian salt gland and teleost gill with formaldehyde as fixative.J. Histochem. Cytochem. 18, 251–63.

    Google Scholar 

  • Evans, A. P. (1969). Reactions of glutaraldehyde and succinaldehyde with compounds containing replaceable hydrogen atoms.Univ. Microfilms Inc. Ann Arbor Michigan 70-11, 634.

    Google Scholar 

  • Fahimi, H. D. &Drochmans (1965a). Essais de standardization de la fixation au glutaraldéhyde I. Purification et détermination de la concentration du glutaraldehyde.J. Microscopie 4, 725–36.

    Google Scholar 

  • Fahimi &Drochmans, P. (1965b). Essais de standardization de la fixation au glutaraldéhyde II. Influence des concentrations en aldéhyde et de l'osmolalité.J. Microscopie 4, 737–48.

    Google Scholar 

  • Fein, M. L. & Harris, E. H. (1962). Quantitative analytical procedure for determining glutaraldehyde and chrome in tanning solutions. U.S. Dept. Agriculture Publication ARS-73-37.

  • Filachione, E. M., Korn, A. H. &Ard, J. S. (1967). The ultraviolet absorption of protein bound glutaraldehyde.J. Am. Leather Chemists Assoc. 67, 450–3.

    Google Scholar 

  • Flitney, E. W. (1966). The time course of fixation of albumin by formaldehyde, glutaraldehyde, acrolein and other higher aldehydes.J. R. microsc. Soc. 85, 353–64.

    Google Scholar 

  • Forssmann, W. G., Siegrist, G., Orci, L., Girardier, L., Pictet, R., Rouiller, C., Baumann, M. &Moritz, A. (1967). Fixation par perfusion pour la microscopie electronique: essai de generalisation.J. Microscopie 6, 279–304.

    Google Scholar 

  • Fortes, P. A. (1971). Glutaraldehyde as a probe of metabolism induced changes in mitochondrial proteins.Proc. Colloq. Johnson Res. Found.

  • Franke, W. W., Krein, S. &Brown, R. M. (1969). Simultaneous glutaraldehyde-osmium tetroxide fixation with post osmication.Histochemie 19, 162–4.

    Google Scholar 

  • French, D. &Edsall, J. T. (1945). The reaction of formaldehyde with amino acids and proteins.Adv. prot. Chem. 2, 277–335.

    Google Scholar 

  • Frigerio, N. A. &Shaw, M. J. (1969). A simple method for determination of glutaraldehyde.J. Histochem. Cytochem. 17, 176–81.

    Google Scholar 

  • Garrett, J. R., Davies, K. J. &Parsons, P. A. (1972). Consumers' guide to glutaraldehyde.Proc. R. microsc. Soc. 7, 116–17.

    Google Scholar 

  • Gil, J. &Weibel, E. R. (1971). An improved apparatus for perfusion.J. Microsc. 94, 241–4.

    Google Scholar 

  • Grantham, J., Cuppage, F. E. &Fanestil (1971). Direct observation of toad bladder response to vasopressin.J. Cell Biol. 48, 695–9.

    Google Scholar 

  • Grillo, T. A. I., Ogunnaire, P. O. &Faoye, S. (1971). Effects of histological and electron microscopical fixatives on the insulin content of rat pancreas.J. Endocr. 51, 645–9.

    Google Scholar 

  • Haas, D. J. (1968). Preliminary studies on the denaturation of crosslinked lysozyme crystals.Biophysical J. 8, 549–55.

    Google Scholar 

  • Habeeb, A. F. S. A. &Hiramoto, R. (1968). Reaction of proteins with glutaraldehyde.Arch. Biochem. Biophys. 126, 16–26.

    Google Scholar 

  • Hanker, J. S., Kasler, F., Bloom, M. G., Copeland, J. S. &Seligman, A. M. (1967). Coordination of polymers of osmium: the nature of osmium black.Science 156, 1737–8.

    Google Scholar 

  • Hardin, J. H. &Spicer, S. S. (1970). Ultrastructure of neuronal nucleoli of rat trigeminal ganglia. Comparison of routine with pyroantimonate-osmium tetroxide fixation.J. Ultrastruct. Res. 31, 16–36.

    Google Scholar 

  • Hardy, P. M., Nicholls, A. C. & Rydon, H. N. (1969). Nature of glutaraldehyde in aqueous solution.Chem. Commun. 565–6.

  • Harries, C. &Tank, L. (1908). Ueber die Aufspaltung der Cyclopentens zum Halbaldehyd der Glutarsaüre bezw. zum Glutardialdehyd.Berlin Ber D. Chem. Ges. 41, 1701–11.

    Google Scholar 

  • Hayat, M. A. (1970).Principles and Techniques of Electron Microscopy. Biological applications. London: Van Nostrand Reinhold Co.

    Google Scholar 

  • Hecker, H. (1970). Ultrastruktur der Symbioten in Ovozyten vonOrnithodorus moubata, Murray (Ixodoidea: Argasidae) nach simultaner Glutaraldehyd-Osmiumfixierung und Nach behandlung mit Uranylacetat (Triple Fixation).Experimentia 26, 874–7.

    Google Scholar 

  • Heller, J., Ostwald, T. J. &Bok, D. (1971). The osmotic behaviour of rod photoreceptor outer segment discs.J. Cell Biol. 48, 633–49.

    Google Scholar 

  • Hirsch, J. G. &Fedorko, M. E. (1968). Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and ‘post fixation’ in uranyl acetate.J. Cell Biol. 38, 615–27.

    Google Scholar 

  • Hodson, S. &Marshall, J. (1967). Tyrosine incorporation into the rabbit retina.J. Cell Biol. 35, 722–6.

    Google Scholar 

  • 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.

    Google Scholar 

  • Hopwood, D. (1967b). The behaviour of various glutaraldehydes on Sephadex G-10 and some implications for fixation.Histochemie 11, 289–95.

    Google Scholar 

  • Hopwood, D. (1967c). The effect of formaldehyde fixation and dehydration on ox adrenal medulla with respect to the chromaffin reaction and post-chroming.Histochemie 10, 98–106.

    Google Scholar 

  • Hopwood, D. (1968). The effect of pH and various fixatives on isolated ox chromaffin granules with respect to the chromaffin reaction.J. Anat. (Lond.)102, 415–24.

    Google Scholar 

  • Hopwood, D. (1969a). Fixatives and fixation: a review.Histochem. J. 1, 323–60.

    Google Scholar 

  • Hopwood, D. (1969b). A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and α-hydroxydipaldehyde with bovine serum albumin and casein.Histochemie 17, 151–61.

    Google Scholar 

  • Hopwood, D. (1969c). Fixation of proteins by osmium tetroxide, potassium dichromate and potassium permanganate.Histochemie 18, 250–60.

    Google Scholar 

  • Hopwood, D. (1970). The reactions between formaldehyde, glutaraldehyde and osmium tetroxide and their fixation effects on bovine serum albumin and on tissue blocks.Histochemie 24, 56–64.

    Google Scholar 

  • Hopwood, D. (1971a). The histochemistry and electron histochemistry of chromaffin tissue.Progr. Histochem. Cytochem. 3, 1–66.

    Google Scholar 

  • Hopwood, D. (1971b). Use of isoelectric focusing to determine the isoelectric point of bovine serum albumin after treatment with various common fixatives.Histochem. J. 3, 201–5.

    Google Scholar 

  • Hopwood, D. (1972a). Fixation with mercury compounds.Progr. Histochem. Cytochem. 4, 193–224.

    Google Scholar 

  • Hopwood, D., Allen, C. R. &Mccabe, M. (1970). The reactions between glutaraldehyde and various proteins. An investigation of their kinetics.Histochem. J. 2, 137–50.

    Google Scholar 

  • Horton, A. A. &Packer, L. (1969). Mitochondrial metabolism of aldehydes.Biochem. J. 116, 19–20.

    Google Scholar 

  • Horton, A. A. &Packer, L. (1969a). Mitochondrial metabolism of aldehydes.Biochem. J. 116, 19P-20P.

    Google Scholar 

  • Horton, A. A. &Packer, L. (1970). Interactions between malondialdehyde and rat liver mitochondria.J. Gerentol 25, 199–204.

    Google Scholar 

  • Hughes, R. C. &Thurman, P. F. (1970). Crosslinking of bacterial cell walls with glutaraldehyde.Biochem. J. 119, 925–6.

    Google Scholar 

  • Jahn, K. (1971). Über histochemische Aldehydreaktionen nach Fixation mit Glutaraldehyd.Acta Histochem. 39, 298–301.

    Google Scholar 

  • Janigan, D. T. (1964). Tissue enzyme fixation studies.Lab. Invest. 13, 1038–50.

    Google Scholar 

  • Janigan, D. T. (1965). The effect of aldehyde fixation on acid phosphatase activity in tissue blocks.J. Histochem. Cytochem. 13, 473–83.

    Google Scholar 

  • Jansen, E. F. &Olson, A. C. (1969). Properties and enzymatic activities of papain insolubilized with glutaraldehyde.Arch. Biochem. Biophys. 129, 221–7.

    Google Scholar 

  • Jansen, E. F., Tomimatsu, Y. &Olsen, A. C. (1971). Cross linking of α-chymotrypsin and other proteins by reaction with glutaraldehyde.Arch. Biochem. Biophys. 144, 394–400.

    Google Scholar 

  • Jansen, E. G., Curl, Al. &Balls, A. K. (1951). A crystalline, active oxidation product of α-chymotrypsin.J. Biol. Chem. 189, 671–82.

    Google Scholar 

  • Jard, S., Bourget, J., Carasso, N. &Favard, P. (1966). Action des fixatives sen la perméabilité et l'ultrastructure de la véssie de grenouille.J. Microscopie 5, 31–50.

    Google Scholar 

  • Jonsson, G. (1971). Quantitation of fluorescence of biogenic amines.Progr. Histochem. Cytochem. 2, 299–334.

    Google Scholar 

  • Kaldor, G. &Weinbach, S. (1965). Use of toluidine diisocyanide and glutaric dialdehyde in the conjugation of heavy meromyosin with light meromyosin.Arch. Biochem. Biophys. 112, 448–52.

    Google Scholar 

  • Kanamura, S. (1970). Difference in resistance to glutaraldehyde or formaldehyde fixation between mouse and rat hepatic glucose 6-phosphatase.Acta Histochem. Cytochem. 3, 160–2.

    Google Scholar 

  • Kanamura, S. (1971). Demonstration of glucose 6-phosphatase activity in hepatocytes following transparenchymal perfusion fixation with glutaraldehyde.J. Histochem. Cytochem. 19, 386–7, 520–1.

    Google Scholar 

  • Karnowsky, M. J. (1967). The ultrastructural basis of capillary permeability. Studies with peroxidase as a trace.J. Cell Biol. 36, 213.

    Google Scholar 

  • Kay, D. (1967).Techniques for Electron Microscopy. 2nd edn. Oxford: Blackwell.

    Google Scholar 

  • Landon, D. N. (1970). The influence of fixation upon the fine structure of the Z-disc of the rat striated muscle.J. Cell Sci. 6, 257–76.

    Google Scholar 

  • 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, 117–21.

    Google Scholar 

  • Levy, W. A., Herzog, I., Suzuki, K., Katzman, R. &Scheinberg, L. (1965). Method for combined ultrastructural and biochemical analysis of normal tissue.J. Cell Biol. 27, 119–32.

    Google Scholar 

  • L'hermite, P. &Israel, M. (1969). Action du glutaraldéhyde sur les lipide et les protéines myéliniques.Ann. Histcchim. 14, 1–11.

    Google Scholar 

  • Lillie, R. D. (1967). Hot chromation oxyphilia.Histochemie 11, 332–49.

    Google Scholar 

  • Ljubešić, N. (1970). Osmiophile substanz in Blattzellen der Brombeere (Rubus fructicosus L.S.L.).Protoplasma 69, 49–59.

    Google Scholar 

  • Mantai, K. E. (1970). Some effects of hydrolytic enzymes on coupled and uncoupled electron flow in chloroplasts.Plant Physiol. 45, 563–6.

    Google Scholar 

  • Marzona, M. &Modica, G. (1969). Stabilization of wool by difunctional aldehydes.Ann. Chim. (Rome) 59, 956–62.

    Google Scholar 

  • Maunsbach, A. B. (1966). The influence of different fixatives and fixation methods on the ultrastructure of rat kidney proximal tubule cells.J. Ultrastruct. Res. 15, 283–309.

    Google Scholar 

  • Milch, R. A. (1963). Studies on collagen tissue aging: interaction of certain intermediary metabolites with collagen.Gercntologia 7, 129–52.

    Google Scholar 

  • Mollenhauer, H. H. &Totten, C. (1971). Studies on seeds I. Fixation of seeds.J. Cell Biol. 48, 387–94.

    Google Scholar 

  • Morel, F. M. M., Baker, R. F. &Wayland, H. (1971). Quantitation of human red blood cell fixation by glutaraldehyde.J. Cell Biol. 48, 91–100.

    Google Scholar 

  • Munton, T. J. &Russell, A. D. (1970). Aspects of the action of glutaraldehyde onEscherichia coli.J. appl. Bact. 33, 410–19.

    Google Scholar 

  • Ogata, K., Ottesen, M. &Svendsen, I. (1968). Preparation of water-insoluble, enzymically active derivatives of subtilisin type Novo by cross linking with glutaraldehyde.Biochem. biophys. Acta 159, 403–5.

    Google Scholar 

  • Ostrowski, K., Komender, J. &Kwarecki, K. (1961). Quantitative investigations of the solubility of proteins extracted from tissues fixed by different chemical and physical methods.Ann. Histochem. 6, 501–6.

    Google Scholar 

  • Ottesen, M. &Svenssen, B. (1971). Modification of papian by treatment with glutaraldehyde under reducing and non-reducing conditions.C.R. Lab. Carlsberg 38, 171–85.

    Google Scholar 

  • Packer, L., Allen, J. M. &Starks, M. (1968a). Light induced transport in glutaraldehyde fixed chloroplasts: studies with nigercin.Arch. Biochem. 128, 142–52.

    Google Scholar 

  • Packer, L. &Greville, G. D. (1969). Energy linked oxidation of glutaraldehyde by rat liver mitochondria.FEBS Lett. 3, 112–14.

    Google Scholar 

  • Packer, L., Wrigglesworth, J. M., Fortes, P. A. G. &Pressman, B. C. (1968b). Expansion of the inner membrane compartment and its relation to mitochondrial volume-ion transport.J. Cell Biol. 39, 382.

    Google Scholar 

  • Papadimitriou, J. M. &Van Duijn, P. (1970). Effects of fixation and substrate protection on the isoenzymes of aspartate aminotransferase studied in a quantitative cytochemical model system.J. Cell Biol. 47, 71–83.

    Google Scholar 

  • Park, R. B., Kelly, J., Drury, S. &Sauer, K. (1966). The Hill reaction of chloroplasts isolated from glutaraldehyde-fixed spinach leaves.Proc. Nat. Acad. Sci. U.S.A. 55, 1056–62.

    Google Scholar 

  • Pearse, A. G. E. (1968).Histochemistry: Theoretical and Applied, 3rd edn., vol. I. London: Churchill.

    Google Scholar 

  • Peters, T. &Ashley, C. A. (1967). An artefact in autoradiography due to binding of free amino acids to tissues by fixatives.J. Cell Biol. 33, 53–60.

    Google Scholar 

  • Quiocho, F. A. &Richards, F. M. (1964). Intermolecular crosslinking of a protein in the crystalline state: carboxypeptidase-A.Proc. Nat. Acad. Sci. U.S.A. 52, 833–8.

    Google Scholar 

  • Quiocho, F. A. &Richards, F. M. (1966). The enzymic behaviour of carboxypeptidase-A in the solid state.Biochem. 5, 4062–76.

    Google Scholar 

  • Rabinovitch, M. &Gary, P. P. (1968). Effect of the uptake of staphylococci on the ingestion of glutaraldehyde treated red cells attached to macrophages.Expt. Cell Res. 52, 363–9.

    Google Scholar 

  • Reale, E. &Luciano, L. (1970). Fixierung mit Aldehyden. Ihre Eignung für histologische und histochemische Untersuchungen in der Licht- und Elecktronen-mikroskopie.Histochemie 23, 144–70.

    Google Scholar 

  • Richards, F. M. &Knowles, J. R. (1968). Glutaraldehyde as a protein cross linking agent.J. Mol. Biol. 37, 231–3.

    Google Scholar 

  • Robertson, E. A. &Schultz, R. L. (1970). The impurities in glutaraldehyde and their effect on the fixation of the brain.J. Ultrastruct. Res. 30, 275–87.

    Google Scholar 

  • Roozemond, R. C. (1969). The effect of fixation with formaldehyde and glutaraldehyde on the composition of phospholipids extractable from rat hypthalamus.J. Histochem. Cytochem. 17, 482–6.

    Google Scholar 

  • Rost, F. W. D. &Ewen, S. W. B. (1971). New methods for the histochemical demonstration of catechol amines, tryptamines, histamine and other arylethylamines by acid- and aldehyde-induced fluorescence.Histochem. J. 3, 207–12.

    Google Scholar 

  • Roux, S. J. &Hillman, W. S. (1969). The effect of glutaraldehyde and two monoaldehydes on phytochrome.Arch. Biochem. Biophys. 131, 423–9.

    Google Scholar 

  • Rubbo, S. D., Gardner, J. F. &Webb, R. L. (1967). Biocidal activities of glutaraldehyde and related compounds.J. appl. Bact. 30, 78–87.

    Google Scholar 

  • Sabatini, D. D., Bensch, K. &Barnnett, R. J. (1963). Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation.J. Cell. Biol. 17, 19–58.

    Google Scholar 

  • Sachs, D. H. &Winn, H. J. (1970). The use of glutaraldehyde as a coupling agent for ribonuclease and bovine serum albumin.Immunochem. 7, 581–5.

    Google Scholar 

  • Schechter, I. (1971). Prolonged survival of glutaraldehyde treated skin homografts.Proc. Nat. Acad. Sci. U.S.A. 68, 1590–3.

    Google Scholar 

  • Schejter, A. &Bareli, A. (1970). Preparation and properties of cross linked water-insoluble catalase.Arch. Biochem. Biophys. 136, 325–30.

    Google Scholar 

  • Schultz, R. L. &Karlsson, U. (1965). Fixation of the central nervous system by aldehyde perfusion II. Effect of osmolarity pH of perfusata and fixative concentration.J. Ultrastruct. Res. 12, 187–206.

    Google Scholar 

  • Schwab, D. W., Janney, A. H., Scala, J. &Lewin, L. M. (1970). Preservation of fine structures in yeast by fixation in a dimethyl sulfoxide-acrolein-glutaraldehyde solution.Stain Technol. 45, 143–7.

    Google Scholar 

  • Seki, M. (1936). Zur physikalischen Chemie der Histologischen Färbung IX. Über den Einfluss der Fixierung auf die Färbarkheit der histologische Element.Z. Zellforsch 18, 21–55.

    Google Scholar 

  • Shea, S. M. (1971). Lanthanum staining of the surface coat of cells.J. Cell Biol. 51, 611–20.

    Google Scholar 

  • Shnitka, T. K. &Seligman, A. M. (1971). Ultrastructural localization of enzymes.Ann. Rev. Biochem. 40, 375–96.

    Google Scholar 

  • Sipes, J. (1970). El uso del glutaraldehido en escala industrial.Rev. Assoc. Argent. Quim. Tech. Indust. Cuer. II, 148–55.

    Google Scholar 

  • Sjöstrand, F. S. &Barajas, L. (1968). Effect of modifications in conformation of protein molecules on structure of mitochondrial membranes.J. Ultrastruct. Res. 25, 121–55.

    Google Scholar 

  • Thiessen, G., Thiessen, H., Dowidat, H. J., Luciano, L. &Reale, E. (1970). Die Diffusion der99Fe-markierten Hämoglobins, ein Artefakt der Glutaraldehyde-Fixierung.Histochemie 23, 171–5.

    Google Scholar 

  • Tomimatsu, Y., Jansen, E. F., Gaffield, W. &Olsen, A. C. (1971). Physical chemical observations on the α-chymotripsin-glutaraldehyde system during formation of an insoluble derivation.J. Colloid Interface Science 36, 51–64.

    Google Scholar 

  • Tooze, J. (1964). Measurements of some cellular changes during the fixation of amphibian erythrocytes w with osmium tetroxide solutions.J. Cell Biol. 22, 551–63.

    Google Scholar 

  • Trelstad, R. L. (1969). Effect of pH on the stability of purified glutaraldehyde.J. Histochem. Cytochem. 17, 756–7.

    Google Scholar 

  • Trump, B. F. &Bulger, R. E. (1966). New ultrastructural characteristics of cells fixed in a glutaraldehyde-osmium tetroxide mixture.Lab. Invest. 15, 368–79.

    Google Scholar 

  • Trump, B. F. &Ericsson, J. L. E. (1965). The effect of the fixative solution on the ultrastructure of cells and tissues. A comparative analysis with particular attention to the proximal convoluted tubule of the rat kidney.Lab. Invest. 14, 1245–325.

    Google Scholar 

  • Vanha-Perttula, T. &Grimley, P. M. (1970). Loss of proteins and other macromolecules during preparation of cell cultures for high resolution autoradiography.J. Histochem. Cytochem. 18, 565–75.

    Google Scholar 

  • Van Harreveld, A. &Steiner, J. (1970). Extracellular space in frozen and ethanol substituted central nervous system.Anat. Rec. 166, 117–30.

    Google Scholar 

  • van Harreveld, A. &Khattab, F. I. (1968). Perfusion fixation with glutaraldehyde and post fixation with osmium tetroxide for electron microscopy.J. Cell Sci. 3, 579–84.

    Google Scholar 

  • Walsh, K. A., Houston, L. L. &Kenner, R. A. (1970).Chemical Modification of Bovine Trypsinogen and Trypsin in Structure Function Relationships of Proteolytic Enzymes (eds. P. Desnuelle, H. Neurath & M. Ottesen). Copenhagen: Munksgaard.

    Google Scholar 

  • Wang, J. H. C. &Tu, J. I. (1969). Modification of glycogen phosphorylase by glutaraldehyde.Biochem. 8, 4403–10.

    Google Scholar 

  • Webster, H. De F. &Ames, A. (1969). Glutaraldehyde fixation of central nervous tissue: an electron microscopic evaluation in the isolated rabbit retina.Tissue Cell 1, 53–62.

    Google Scholar 

  • Webster, H. De F. &Ames, A. &Nesbett, F. B. (1969). A quantitative morphological study of osmotically induced swelling and shrinkage in nervous tissue.Tissue Cell 1, 201–6.

    Google Scholar 

  • West, J. &Mangan, J. L. (1970). Effects of glutaraldehyde on the protein loss and photochemical properties of kale chloroplasts: preliminary studies on food conversion.Nature (Lond.) 228, 466–8.

    Google Scholar 

  • Wrigglesworth, J. M., Packer, L. &Branton, D. (1970). Organization of mitochondrial structure as revealed by freeze etching.Biochem. Biophys. Acta 205, 125–35.

    Google Scholar 

  • Yokota, K., Suzuki, Y. & Ishii, Y. (1965). Temperature dependence of the polymerization modes of glutaraldehyde. In:Chem. Abstr. 65, 13835.

  • Young, E. G. (1963).Comprehensive Biochemistry. Vol. 7, p. 25. New York: Elsevier.

    Google Scholar 

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Hopwood, D. Theoretical and practical aspects of glutaraldehyde fixation. Histochem J 4, 267–303 (1972). https://doi.org/10.1007/BF01005005

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