Synopsis
Frog sartorius muscles have been fixed sequentially with acrolein and osmium tetroxide dissolved in vehicles of various tonicities, and the myosin filament spacings and sarcomere lengths measured with the electron microscope. From these dimensions the myosin unit-cell volume has been calculated and compared with X-ray diffraction data to determine the effect of fixation. In muscles soaked in normal Ringer and afterwards fixed using normal Ringer as a vehicle for the fixation agents, the unit-cell volume undergoes a 10.4% reduction during the preparative procedure. Muscles soaked in hypertonic Ringer undergo a similar reduction in volume during fixation, provided hypertonic Ringer is used as the vehicle; if they are fixed in normal Ringer, the lattice swells during fixation, even if the change to the normal tonicity vehicle occurs after acrolein fixation. If blocks suitable for embedding are cut from the muscles before, rather than after, osmium fixation, more complex changes in intracellular dimensions may occur, including artefactual swelling of the T-system. It is concluded that fixation of tissues exposed to modifications of normal physiological solutions should be performed using the same modified solutions as fixative vehicles.
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
APRIL, E. W., BRANDT, P. W. & ELLIOT, G. F. (1971). The myofilament lattice: studies on isolated fibers. I. The constancy of the unit-cell volume with variation in sarcomere length in a lattice in which the thick-to-thin myofilament ratio is 6:1. J. Cell. Biol.51, 72–82.
BIRKS R. I. (1971). Effects of stimulation on synaptic vesicles in sympathetic ganglia, as shown by fixation in the presence of Mg2+. J. Physiol., Lond. 216, 26–28P.
BIRKS, R.I. & DAVEY, D.F. (1969). Osmotic responses demonstrating the extracellular character of the sarcoplasmic reticulum. J. Physiol., Lond.202, 171–88.
BIRKS, R. I. & DAVEY, D. F. (1972). An analysis of volume changes in the T-tubes of frog skeletal muscle exposed to sucrose. J. Physiol., Lond.222, 95–111.
BONE, Q. & DENTON, E.J. (1971). The osmotic effects of electron microscope fixatives. J. Cell Biol.49, 571–81.
BONE, Q. & RYAN, K. P. (1972). Osmolarity of osmium tetroxide and glutaraldehyde fixatives. Histochem. J.4, 331–47.
BRANDT, P. W., LOPEZ, E., RUEBEN, J. P. & GRUNDFEST, H. (1967). The relationship between myofilament packing density and sarcomere length in frog striated muscle. J. Cell Biol.33, 255–63.
BRANDT, P. W., REUBEN, J. P. & GRUNDFEST, H. (1968). Correlated morphological and physiological studies on isolated single muscle fibers. II. The properties of the crayfish transverse tubular system: localization of the sites of reversible swelling. J. Cell Biol.38, 115–29.
CARSLEN, F., KNAPPEIS, G. G. & BUCHTAL, F. (1961). Ultrastructure of the resting and contracted striated muscle fibers at different degrees of stretch. J. biophys. Cytol., 11, 95–117.
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, 529–33.
ELBERS, P. F. (1966). Ion permeability of the egg of Limnaea stagnalis L., on fixation for electron microscopy. Biochim. biophys. Acta 112, 318–29.
FOULKS, J. G., PACEY, J. A. & PERRY, FLORENCE A. (1965). Contractures and swelling of the transverse tubules during chloride withdrawal in frog skeletal muscle. J. Physiol., Lond.180, 96–115.
FREYGANG, W. H., GOLDSTEIN, D. A., HELLAM, D. C. & PEACHEY, L. D. (1964). The relation between the late after-potential and the size of the transverse tubular system of frog muscle. J. gen. Physiol.48, 235–63.
FREYGANG, W. H., RAPOPORT, S. I. & PEACHEY, L. D. (1967). Some relations between changes in the linear electrical properties of striated muscle fibers and changes in ultrastructure. J. gen. Physiol.50, 2437–58.
GOOD, N.E., WINGET, G.D., WINTER, WILHELMINA, CONNOLLY, T. N., IZAWA, S. & SINGH, R. M. M. (1966). Hydrogen ion buffers for biological research. Biochemistry, N.Y.5, 467–77.
GRIFFIN, J. L. (1963). Motion picture analysis of fixation for electron microscopy: Amoeba proteus. J. Cell Biol.19, 77A.
HAYAT, M. A. (1970). Principles and Techniques of Electron Microscopy. Volume I, Biological applications. New York: Van Nostrand Reinhold.
HUXLEY, H. E. (1957). The double array of filaments in cross-striated muscle. J. biophys. biochem. Cytol.3, 631–48.
HUXLEY, H. E. (1968). Structural difference between resting and rigor muscle: evidence from intensity changes in the low-angle equatorial X-ray diagram. J. molec. Biol.37, 507–20.
HUXLEY, H. E., PAGE, SALLY & WILKIE, D. R. (1963). An electron microscopic study of muscle in hypertonic solutions. Appendix to Dydyhska, Maria & Wilkie, D. R. J. Physiol., Lond.169, 312–29.
KARNOVSKY, M. J. (1967). The ultrastructural basis of capillary permeability studied with peroxidase as a tracer. J. Cell Biol.35, 213–36.
KELLENBERGER, E., RYTER, ANTOINETTE & SÉCHAUD, JANINE (1958). Electron microscope study of DNA-containing plasms. II. Vegetative and mature phage DNA as compared with normal bacterial nucleoids in different physiological states. J. biophys. biochem. Cytol.4, 671–8.
LUFT, J. H. (1959). The use of acrolein as a fixative for light and electron microscopy. Anat. Rec.133, 305.
MILLONIG, G. (1968). Experimental approach to problems of fixation and dehydration. Advances in Optical and Electron Microscopy 2, 287–317.
PAGE, SALLY G. & HUXLEY, H. E. (1963). Filament lengths in striated muscle. J. Cell Biol.19, 369–90.
PALADE, G. E. (1952). A study of fixation for electron microscopy. J. exp. Med.95, 285–98.
PEACHEY, L. D. (1958). Thin sections. I. A study of section thickness and physical distortion produced during microtomy. J. biophys. biochem. Cytol.4, 233–42.
RAPOPORT, S. I., PEACHEY, L.D. & GOLDSTEIN, D. A. (1969). Swelling of the transverse tubular system in frog sartorius. J. gen. Physiol.54, 166–77.
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.
ROME, ELIZABETH (1968). X-ray diffraction studies of the filament lattice of striated muscle in various bathing media. J. molec. Biol.37, 331–44.
SPURR, A. R. (1969). A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res.26, 31–43.
WOOD, R. L. & LUFT, J. H. (1965). The influence of buffer systems on fixation with osmium tetroxide. J. Ulstrastruct. Res.12, 22–45.
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
Davey, D.F. (1973). The effect of fixative tonicity on the myosin filament lattice volume of frog muscle fixed following exposure to normal or hypertonic Ringer. In: Stoward, P.J. (eds) Fixation in Histochemistry. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-3260-0_4
Download citation
DOI: https://doi.org/10.1007/978-1-4899-3260-0_4
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-412-12050-3
Online ISBN: 978-1-4899-3260-0
eBook Packages: Springer Book Archive