Examination and Evaluation
in the Electron Microscope
Sorry, but you are on your own with this part. We haven't come far enough yet to be able to teach this important part of microscopy through this medium. However, books still exist. Here is a list of some which may help interpret EM data. A short list of reference books and useful journal references is also included.
Books on ultrastructure and immunocytochemistry
Fawcett, D. W. 1981. The Cell. W. B. Saunders Company. Philadelphia.
Griffiths, G. 1993. Fine Structure Immunocytochemistry. Springer Verlag, Heidelberg.
Larsson, L. I. 1988. Immunocytochemistry: theory and practice. CRC Press, Boca Raton, Florida.
Bozzola, J. J. and L. D. Russell. 1992. Electron Microscopy. Jones and Bartlett, Boston.
Reference books on transmission electron microscopy
Dawes, C. J. 1979. Biological techniques in electron microscopy. Barnes and Noble, New York.
Glauert, A. M. (editor) Practical methods in electron microscopy. North Holland, Amsterdam.
Volume 1: Glauert, A. M. 1974. Fixation, dehydration and embedding of biological specimens.
Volume 2: Agar, A. W., R. H. Anderson and D. Chescoe. 1974. Principles and practice of electron microscope operation.
Volume 5: Lewis, P. R. and D. P. Knight. 1977. Staining methods for sectioned material.
Hayat, M. A. 1981. Fixation for electron microscopy. Academic Press, New York.
Hayat, M. A. 1986. Basic techniques for transmission electron microscopy. Academic Press, New York.
Hayat, M. A. (editor) Correlative Microscopy in biology: Instrumentation and methods. Academic Press, New York. Weakley, B. S. 1981. A beginner's handbook in biological electron microscopy. Churchill Livingstone, London, New York.
Useful references in scientific journals for immunocytochemical methods.
Liou, W., Geuze H.J. & Slot J.W. 1996. Improving structural integrity of cryosections for immunogold labeling. Histochem. Cell Biolo. 106:41-58.
Useful review discussing methods of improving specimen morphology using modified sectioning and section retrieval techniques.
Oprins, A. Geuze H.J. & Slot J.W. 1994. Cryosubstitution dehydration of aldehyde-fixed tissue: a favorable approach to quantitative immunocytochemistry. J. Histochem. Cytochem. 42:497-503.
Examination of quantitation using fixed, cryoprotected and frozen samples embedded in Lowicryl.
van Genderen, I.L. van Meer G. Slot J.W. Geuze H.J. & Voorhout W.F. 1991. Subcellular localization of Forssman glycolipid in epithelial MDCK cells by immuno-electronmicroscopy after freeze-substitution. J. Cell Biol. 115:1009-1019.
Description of effect different embedding methods have on localization of potentially soluble antigens within cells.
Slot, J.W. Geuze H.J. Gigengack S. Lienhard G.E. & James D.E. 1991 Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J. Cell Biol. 113:123-135.
Method for blocking protein A-gold cross-reactivity when using two protein A-binding antibodies to sequentially double label cryosections.
Slot, J.W. Posthuma G. Chang L.Y. Crapo J.D. & Geuze H.J. 1989. Quantitative aspects of immunogold labeling in embedded and in nonembedded sections. Am. J. Anat. 185:271-281.
Slot, J.W. & Geuze H.J. 1985. A new method of preparing gold probes for multiple-labeling cytochemistry. Europ. J. Cell Biol. 38:87-93.
Use of tannic acid and sodium citrate to reduce gold chloride and produce colloidal gold with homogeneous particle size.
Geuze, H.J. Slot J.W. van der Ley P.A. & Scheffer R.C. 1981. Use of colloidal gold particles in double-labeling immunoelectron microscopy of ultrathin frozen tissue sections. J. Cell Biol. 89:653-665.
Description of how to apply multiple antibody and protein A-gold reagents with suitable controls.
Slot, J.W. & Geuze H.J. 1981. Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J. Cell Biol. 90:533-536.
Describes the use of sucrose gradients to produce colloidal gold particles with homogeneous size range.
Webster, P. 1998. The production of cryosections through fixed and cryoprotected biological material and their use in immunocytochemistry. Methods in Molecular Biology. vol 117 ch. 4. (Ed. N. Hajibagheri).
Detailed description on how to cryosection using the Tokyuasu method.
