Summary
Nitrogen mustard N-oxide was tried for the fixation of tissue for electron microscopy. A fixative consisting of 1% nitrogen mustard N-oxide, 1% glutaraldehyde and 1% paraformaldehyde buffered at pH 7.4 followed by 1% OsO4 buffered at pH 7.4 was found useful for the tissues examined: thyroid, anterior pituitary, adrenal gland and oviduct of mice.
If the tissues are fixed and the sections are stained with uranyl acetate and lead acetate doubly, the follicle colloid, colloid droplets, and secretory granules containing thyroglobulin in the thyroid become higher in electron density. The cisterna of the maturing face of the Golgi apparatus, secretory granules, ribosomes, nucleolus and chromatin in the cells examined are extremely electron dense. Tubular elements of smooth endoplasmic reticulum in the adrenal cortical cell and microtubules in all the cells examined are also well preserved. The fixative containing nitrogen mustard N-oxide is useful also for cytochemistry. Using tissue fixed by this method and stained en bloc by uranyl acetate, the noradrenaline and adrenaline cells in the adrenal medulla are clearly distinguished by light microscopy.
Similar content being viewed by others
References
Bergmann, M., Stahmann, M.A.: Chemical reactions of nitrogen mustard gasses. VIII. The oxidation of the nitrogen mustard gasses by peracids. J. Org. Chem. 11, 586–591 (1946)
Hayat, M.A.: Principles and techniques of electron microscopy. Biological applications. London: Van Nostrand Reinhold, 1970
Hopwood, D.: Glutaraldehyde fixation. In: Fixation in histochemistry. (ed. by P.J. Stoward). London: Chapman and Hall, 1973
Ishidate, M., Kobayashi, K., Sakurai, Y., Sato, H., Yoshida, T.: Experimental studies on chemotherapy of malignant growth employing Yoshida sarcoma animals. II. The effect of N-oxide derivatives of nitrogen mustard. Proc. Jap. Acad. Sci. 27, 493–500 (1951)
Ross, W.C.J.: In vitro reactions of biological alkylating agents. Ann. N.Y. Acad. Sci. 68, 669–681 (1958)
Author information
Authors and Affiliations
Additional information
This study was supported by a grant from the Japan Educational Ministry
Rights and permissions
About this article
Cite this article
Fujita, H., Komatsu, M. & Nakajima, T. Use of nitrogen mustard N-oxide for the fixation of electron microscopic tissues. Histochemistry 58, 49–56 (1978). https://doi.org/10.1007/BF00489948
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00489948