Summary
For studies on cathodoluminescence, we equipped a scanning electron microscope with a prism spectrometer and sensitive photomultiplier. The apparatus is described and our initial results are presented on the analyse of cathodoluminescence.
The material observed primarily involved studies of immunofluorescent specimens. Human lymphocytes were labelled with a fluorescent antibody and cryosections of rat kidney with Masugi nephritis were labelled with a fluorescent specific antibody. Our apparatus permitted monochromatic imaging of cathodoluminescence emissions and resulted in much improved micrographs. Some possible improvements of the technique are discussed.
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References
Avrameas, J., Ternynck, T.: Peroxidase labelled antibody and Fab conjugated with enhanced intracellular penetration. Immunochem. 8, 1175–1179 (1971)
Barnett, W.A., Wise, M.L.H., Jones, E.C.: Cathodoluminescence of biological molecules, macromolecules and cells. J. Microscopy 105, 299–303 (1975)
Bernard, R., Davoine, F., Pinard, P.: Extension de la technique de microscopie par balayage à l'observation de substances fluorescentes. C.R. Acad. Sci. (Paris) 248, 2564–2566 (1959)
Bond, E.F., Beresford, D., Haggis, G.H.: Improved cathodoluminescence microscopy. J. Microscopy 100, 271–282 (1974)
Carteaud, J.P., Muir, M.D., Grant, P.R.: Pityriasis versicolor: emissive and cathdoluminescence examination in the scanning electron microscope. Sabouraudia 10, 143–146 (1970)
Cavellier, J.F.: Sur l'utilisation de la cathodoluminescence en biologie. J. Microscopie Biol. Cell. 22, 371–380 (1975)
Coons, A.A., Creech, H.J., Jones, R.W.: Immunological properties of an antibody containing a fluorescent group. Proc. Soc. Expl. Biol. Med. 47, 200–202 (1941)
De Mets, M., Lagasse, A.: an investigation of some organic chemicals as cathodoluminescentedyes using the SEM. J. Microscopy 94, 151–156 (1971)
Giles, P.L.: Cathodoluminescence. J. Microscopy Biol. Cell. 22, 357–370 (1975)
Haggis, G.H., Bond, E.F., Pulcher, R.G.: Improved resolution in cathodoluminescent microscopy of biological material. J. Microscopy 108, 177–184 (1976)
Hanusiak, W.M., White, E.W.: SEM cathodoluminescence for characterization of damaged and undamaged α quartz in respirable dusts. Proc. of the 8th annual scanning electron microscope symposium, p. 125–132 III Res. Inst., 125–132, Chicago: III Research Institute
Hartmann, L., Bariety, J., Nebut, M., Lagrue, G.: Masugi hetero-immune nephritis in rat histological and ultrastructural study (attempt of comparison with prolin data). Ann. Biol. Clin. 27, 437–450 (1969)
Holt, D.B.: Quantitative scanning electron microscope studies of cathodoluminescence in Adamantine semi-conductors. In: Quantitative scanning electron microscopy (ed. D.B. Holt, M.D. Muir, P.R. Grant., J.M. Boswarva) p. 335–386. London-New York, San Francisco: Academic Press 1974
Hörl, E.M.: Scanning electron microscopy of biological material using cathodoluminescence. Micron 3, 540–544 (1972)
Hörl, E.M., Buschbeck, F.: Colour scanning electron microscopy. Proc. of the 6th European Congress on Electron Microscopy (ed. D.G. Brandon) Vol. I, p. 331–333 (1976)
Judge, F.J., Stubb, J.M., Philp, J.: A concave mirror, light pipe photon collecting system for cathodoluminescent studies on biological specimens in the J SM 2 scanning electron microscope. J. Physics, E: Scient. Instr. 7, 173–174 (1974)
Manger, W.M., Bessis, M.: Cathodoluminescence produced in cells and proteins by paraformaldehyde as seen with the scanning electron microscope. Proc. of 7th International Congress on electron microscopy, (ed. P. Favard) Vol. I, p. 483. Grenoble 1970
Muir, M.D., Grant, P.R., Hubbard, G., Mundell, J.: Cathodoluminescence spectra. Proc. of the 4th Annual scanning electron microscope symposium, p. 401–408. Chicago: IIII Research Institute 1971
Nakane, P.K., Pierce, G.B.: Enzyme labelled antibodies for the light and electron microscopic localisation of tissue antigens. J. Cell Biol. 33, 307–318 (1967)
Pease, R.F.W., Hayes, T.L.: Scanning electron microscopy of biological material. Nature 210, 1049 (1966)
Remond, G., Kimoto, S., Okuzumi, H.: Use of the SEM in cathodoluminescence observations in natural samples. Proceed. of the 3rd annual scanning electron microscope symposium, p. 34–39. Chicago: III Research Institute 1970
Remond, G.: Applications of cathodoluminescence in mineralogy. J. luminescence 15, 121–155 (1977)
Schmidt, E.H., Brocker, W., Wagner, H., Pfefferkorn, G., Beller, F.K.: Glomerular microcapillary thrombosis demonstrated by the new technique of immunocathodoluminescence. Am. J. Pathology 81, 43–46 (1975)
Semo, J.: Spectroscopie optique en cathodoluminescence au microscope électronique à balayage. Rev. Phys. Appl. 9, 355–359 (1974)
Semo, J.: Etude et réalisation d'un spectroscope pour cathodoluminescence en microscopie électronique à balayage et applications aux semi-conducteurs à large bande interdite. Thèse de troisième cycle, Université de Paris VI (1975)
Steyn, J.B., Giles, P.L., Holt, D.B.: An efficient spectroscopic detection system for cathodoluminescence mode scanning electron microscopy. J. Microscopy 107, 107–128 (1976)
Williams, P.M., Yoffe, A.D.: Monochromatic cathodoluminescence image in the scanning electron microscope. Nature 221, 952–953 (1969)
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Cavellier, J.F., Berry, J.P. & Lagrue, G. Cathodoluminescence applied to immunofluorescence: Present state and improved technical prospects by prism spectrometer light selection. Histochemistry 57, 313–322 (1978). https://doi.org/10.1007/BF00492667
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DOI: https://doi.org/10.1007/BF00492667