Synopsis
The evidence available to date for the importance of fixative osmolarity is considered together with some observations on the volume changes of crab axons after fixation by osmium tetroxide and glutaraldehyde. The results obtained are compared with those obtained from crab axons and from amphioxus skin cells which had been processed and examined with the electron microscope after initial fixation in fixatives of different composition. It is concluded that the osmolarity of the fixative vehicle is of considerable importance when the fixing agent is glutaraldehyde but is of less importance when the fixing agent is osmium tetroxide or a mixture of the two agents. Preliminary observations upon crab axons fixed with glutaraldehyde in a vehicle approximating to the internal composition of the cells suggest that this approach to the design of fixative vehicles may be useful.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
ABBOTT, B. C., HILL, A. V. & HOWARTH, J. V. (1958). The positive and negative heat production associated with a nerve impulse. Proc. Roy. Soc. Lond. B 148, 149–87.
BAHR, G. F., BLOOM, G. & FRIBERG, U. (1957). Volume changes of tissue in physiological fluids during fixation in osmium tetroxide or formaldehyde and during subsequent treatment. Exp. Cell Res. 12, 342–55.
BAKER, J. R. (1933). Cytological technique. London: Methuen.
BAKER, J. R. (1958). Principles of biological microtechnique. London: Methuen.
BAKER, J. R. (1965). The fine structure produced in cells by fixatives. J. Roy. micr. Soc. 84, 115–31.
BIRKS, R. I. & DAVEY, P. F. (1972). An analysis of volume changes in the T-tubes of frog skeletal muscle exposed to sucrose. J. Physiol., Lond. 222, 95–111.
BRINLEY, F. J. (1965). Sodium, potassium and chloride concentrations and fluxes in the isolated giant axon of Homarus. J. Neurophysiol. 28, 742–72.
BOHMAN, S-O. & MAUNSBACH, A. B. (1970). Effects on tissue fine structure of variations in colloid osmotic pressure of glutaraldehyde fixative. J. Ultrastruct. Res. 30, 195–208.
BONE, Q. & DENTON, E. J. (1971). The osmotic effects of electron microscope fixatives. J. Cell Biol. 49, 571–81
BUSSON-MABILLOT, S. (1971). Influence de 1a fixation chimique sur 1es ultrastructures. I. Etude sur 1es organites du follicule Ovarien d’un poisson teleosteen. J. Microsc. 12, 317–48.
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.
CRAWFORD, G. N. C. & BARER, R. (1951). The action of formaldehyde on living cells as studied by phase-contrast microscopy. Quart. J. micr. Sci. 92, 403–52.
CHUANG, S. H. (1968). Seawater and osmium tetroxide fixation of marine animals. In: Cell structure and its interpretation (eds. S. M. McGee Russell & K. F. A. Ross). St Martin’s Press: New York.
DEL CERRO, M. P. & SNIDER, R. S. (1972). Studies on the developing cerebellum. II. The ultra- structure of the external granular layer. J. comp. Neurol. 144, 131–64.
DOGGENWEILER, C. F. & HEUSER, J. E. (1967). Ultrastructure of the prawn nerve sheaths. Role of fixative and osmotic pressure in vesiculation of thin cytoplasmic laminae. J. Cell Biol. 34, 407–20.
ELBERS, P. F. (1966). Ion permeability of the eggs of Limnaea stagnalis L. on fixation for electron microscopy. Biochim. biophys. Acta 112, 318–19.
ELFORD, B. C. & WALTER, C. A. (1972). Preservation of structure and function of smooth muscle. cooled to — 79°C in unfrozen aqueous media. Nature, Lond. 236, 58–9.
HERTWIG, G. (1931). Der Einfluss der Fixierung auf das Kern- und Zell-volumen. Z. Mikr. anat. Forsch 23, 484–504.
HILL, D. K. (1950). The volume change resulting from stimulation of a giant nerve fibre. J. Physiol, Lond. ii, 304–27.
HOPWOOD, D. (1970). The reactions between formaldehyde, glutaraldehyde and osmium tetrox- ide, and their fixation effects on bovine serum albumin and on tissue blocks. Histochemie 24, 50–64.
KARNOVSKY, M. J. (1965). A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J. Cell Biol. 27, 137A.
LUCKE, B. (1940). The living cell as an osmotic system and its permeability to water. In: Cold Spring Harbor Symp. Quant. Biol. 8, 123–32.
MALHOTRA, S. K. (1962). Experiments on fixation for electron microscopy. I. Unbuffered osmium tetroxide. Quart. J. micr. Sci. 103, 5–15.
MAUNSBACH, A. B. (1966). The influence of different fixatives and fixation methods on the ultra-structure of rat kidney proximal tubule cells. II. Effect of varying osmolality, ionic strength, buffer system and fixative concentration of glutaraldehyde solution. J. Ultrastruct. Res. 15, 283–309.
MILLONIG, G. (1968). In Millonig, G. and Marinozzi, V. Fixation and embedding in electron microscopy. In: Advances in optical and electron microscopy (eds. V. E. Coslett & R. Barer). Vol. 2, P. 251. Academic Press: New York.
OLSSON, R. (1961). The skin of amphioxus. Z. Zeilforsch. 64, 90–104.
OVALLE, W. K. (1970). Fine structure of rat intrafusal muscle fibres. The polar region. J. Cell Biol. 51, 83.
PERRACHIA, C. (1970). A system of parallel septa in crayfish nerve fibers. J. Cell Biol. 44, 125–33.
PONDER, E. (1940). The red cell as an osmometer. In: Cold Spring Harbor Symp. Quant. Biol. 8, 133–43.
REYNOLDS, E. S. (1963). The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–12.
ROBERTSON, E. A. & Schultz, R. L. (1970). The impurities in commercial glutaraldehyde and their effect on the fixation of brain. J. Ultrastruct. Res. 30, 275–87.
SJöSTRAND, F. S. (1956). Electron microscopy of cells and tissues. In: Physical Techniques in Biological Research (eds. G. Oster & A. W. Pollister). Vol. 3, pp. 241–97. Academic Press: New York.
TILNEY, L. G. & GODDARD, J. (1970). Nucleating sites for the assembly of cytoplasmic microtubules in the ectodermal cells of blastulae of Arbacia punctulata. J. Cell Biol. 46, 564–75.
YOUNG, J. Z. (1935). Osmotic pressure of fixing solutions. Nature, Lond. 135, 823–5.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1973 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Bone, Q., Ryan, K.P. (1973). Osmolarity of osmium tetroxide and glutaraldehyde fixatives. In: Stoward, P.J. (eds) Fixation in Histochemistry. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-3260-0_3
Download citation
DOI: https://doi.org/10.1007/978-1-4899-3260-0_3
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-412-12050-3
Online ISBN: 978-1-4899-3260-0
eBook Packages: Springer Book Archive