Summary
T-lymphocytes derived from human peripheral blood and passed through a nylon-wool column, were employed to develop and test a new Stereological model system for free spherical cells, allowing a quantitative characterization of the cell and its components at the ultrastructural level. Electron micrographs were recorded in a hierarchical manner at three different levels of magnification and subjected to point counting procedures. The resulting parameters were expressed in relation to various reference compartments, both absolute and relative. Results indicated that the average volume of a small, non-activated T-lymphocyte was 103.8 μm3, the nuclear volume 47.5 μm3 and the cytoplasmic volume 55.9 μm3. On the average, the cytoplasm contained 30 mitochondria, 0.7 μm3 RER-cisternae, 0.2 μm3 cisternae and vesicles of the Golgi apparatus and about 231,000 free ribosomes (most of them single). The ratio of eu- to heterochromatin volume was 0.5. The design and application of the Stereological model system are discussed with regard to dynamic studies of a variety of free cells, such as macrophages, neutrophilic granulocytes and various lymphocytes.
Zusammenfassung
Menschliche, in Nylonwolle gereinigte T-Lymphozyten aus dem peripheren Blut dienten als repräsentatives Untersuchungsobjekt zur Schaffung eines neuen stereologischen Modellsystems für freie, sphärische Zellen. Dieses System erlaubt, die Zelle und die darin enthaltenen Strukturkomponenten auf ultrastruktureller Ebene quantitativ zu charakterisieren.
Similar content being viewed by others
References
Bennett, H.S., Luft, J.H.: S-collidine as a basis for buffering fixatives. J. biophys. biochem. Cytol. 6, 113–114 (1959)
Biberfeld, P.: Morphogenesis in blood lymphocytes stimulated with phytohaemagglutinin (PHA). A light and electron microscopic study. Acta path, microbiol. scand., Suppl. 223 (1971)
Böyum, A.: Isolation of mononuclear cells and granulocytes from human blood. Scand. J. clin. Lab. Invest. 21 (Suppl. 97), 77–89 (1968)
Burckhardt, J.J., Guggisberg, E., Fellenberg, R. von: Thymus immunglobulin receptors. Immunology 26, 521–537 (1974)
Burri, P.H., Giger, H., Gnägi, H.R., Weibel, E.R.: Application of stereologic methods to cytophysiologic experiments on polarized cells. In: Electron microscopy, Vol. 1 (D.S. Bocciarelli, ed.), p. 593, IV. European Conference, Rome (1968)
DeHoff, R.T., Rhines, F.N.: Determination of the number of particles per unit volume from measurements made on random plane sections: the general cylinder and the ellipsoid. Trans. AIME 221, 975–982 (1961)
Douglas, St.D., Cohnen, G., Brittinger, G.: Ultrastructural comparison between phytomitogen transformed normal and chronic lymphocytic leukemia lymphocytes. J. Ultrastruct. Res. 44, 11–26 (1973)
Fossum, S., Gautvik, K.M.: Stereological and biochemical analysis of prolactin and growth hormone secreting rat pituitary cells in culture. Cell Tiss. Res. 184, 169–178 (1977)
Fraska, J.M., Parks, V.R.: A routine technique for double staining ultrathin sections using uranyl and lead salts. J. Cell Biol. 25, 157–161 (1965)
Giger, H., Riedwyl, H.: Bestimmung der Größenverteilung von Kugeln aus Schnittkreisradien. Biometr. Z. 12, 156–162 (1970)
Glagoleff, A.A.: On the geometrical methods of quantitative mineralogic analysis of rocks. Trans. Inst. Econ. Min. and Metal. Moscow 59, 475 (1933)
Greaves, M.F., Brown, G.: Purification of human T- and B-lymphocytes. J. Immunol. 112, 420–423 (1974)
Heiniger, H.J., Riedwyl, H., Giger, H., Sordat, B., Cottier, H.: Ultrastructural differences between thymic and lymph node small lymphocytes of mice. Nucleolar size and cytoplasmic volume. Blood 30, 288–300 (1967)
Hirsch, J.G., Fedorko, M.E.: Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and “postfixation” in uranyl acetate. J. Cell Biol. 38, 615–627 (1968)
Hirschhorn, R., Decsy, M.I., Troll, W.: The effect of PHA stimulation of human peripheral blood lymphocytes upon cellular content of euchromatin and heterochromatin. Cell Immunol. 2, 696–701 (1971)
Hoffstein, S., Zurier, R.B., Weissmann, G.: Mechanism of lysosomal enzyme release from human leukocytes. III. Quantitative morphologic evidence for an effect of cyclic nucleotides and colchicine on degranulation. Clin. Immunol. Immunopath. 3, 201–217 (1974)
Huhn, D., Rodt, H., Thiel, E., Fink, U., Ruppelt, W.: Elektronenmikroskopische Untersuchungen an menschlichen Lymphozyten. Blut 32, 87–102 (1976)
Karnovsky, M.J.: A formaldehyde glutaraldehyde fixative of high osmolarity for use in electron microscopy. J. Cell Biol. 27, 173A-138A (1965)
Konwinski, M., Kozlowski, T.: Morphometric study of normal and phytohemagglutinin-stimulated lymphocytes. Z. Zellforsch. 129, 500–507 (1972)
Le Bouteiller, Ph., Kinsky, R.G., Vujanovic, N., Duc, H.T., Voisin, G.A.: Morphological differences between thymus and bone-marrow-derived lymphocytes. II. An electron microscopic and experimental study in unstimulated mice. Differentiation 6, 125–141 (1976)
Levy, J.D., Knieser, M.R., Briggs, W.A.: Ultrastructural characteristics of the non-immune rosette- forming cell. J. Cell Sci. 18, 79–96 (1975)
Luft, J.H.: Improvements in epoxy-resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961)
Matter, A., Lisowska-Bernstein, B., Ryser, J.E., Lamelin, J.P., Vasalli, P.: Mouse thymus-independent and thymus-derived lymphoid cells. II. Ultrastructural studies. J. exp. Med. 136, 1008–1030 (1972)
Mayhew, T.M., Williams, M.A.: A comparison of two sampling procedures for stereologic analysis of cell pellets. J. Microscopy (Lond.) 94, 195–204 (1971)
Mayhew, T.M., Williams, M.A.: A quantitative morphological analysis of macrophage stimulation. I. A study of subcellular compartments and of the cell surface. Z. Zellforsch. 147, 567–588 (1974)
Muñiz, F.J., Houston, E.W., Cruz-Abad, L., Ritzmann, S.E., Levin, W.C.: Nuclear volume distribution of PHA-stimulated human lymphocytes. Proc. Soc. exp. Biol. (N.Y.) 135, 334–339 (1970)
Preud'homme, J.L., Labaume, S.: Immunofluorescent staining of human lymphocytes for the detection of surface immunoglobulins. In: Fifth International Conference on Immunofluorescence and Related Staining Techniques. Ann. N.Y. Acad. Sci. 254, 254–261 (1975)
Reynolds, E.S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963)
Schwenk, H.U., Gimpert, E., Plüss, H.J., Pilgrim, U., Hitzig, W.H.: Lymphocyte markers for B- und T-cells in primary immunodeficiency diseases. Klin. Wschr. 52, 426–432 (1974)
Simar, L.J..: L'ultrastructure des ganglions lymphatiques au cours de la réponse immunitaire. Thèse. Liège, Belgium (1973)
Sipe, C.R., Chanana, A.D., Chronkite, E.P., Gulliani, G.L., Joel, D.D.: Studies on lymphocytes. XIII. Nuclear volume measurement as a rapid approach to estimate the proliferative fraction. Scand. J. Haemat. 16, 196–201 (1976)
Sören, L., Biberfeld, P.: Quantitative studies on RNA accumulation in human PHA-stimulated lymphocytes during blast transformation. Exp. Cell Res. 79, 359–367 (1973)
Stewart, C.C., Cramer, St.F., Steward, P.G.: The response of human peripheral blood lymphocytes to phytohemagglutinin: Determination of cell numbers. Cell Immunol. 16, 237–250 (1975)
Valkov, I., Moyne, G., Robineaux, R.: Etude morphométrique de certaines structures nucléoprotéiques des noyaux de lymphocytes cultivés in vitro en présence de phytohémagglutinine. Europ. J. Immunol. 4, 570–577 (1974)
Weibel, E.R.: Automatic sampling stage microscope and data printout unit. In stereology (H. Elias, ed.). Berlin-Heidelberg-New York: Springer 1967
Weibel, E.R.: Stereologic principles for morphometry in electron microscopic cytology. Int. Rev. Cytol. 26, 235–302 (1969)
Weibel, E.R., Bolender, R.P.: Stereologic techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy, Vol. 3 (M.A. Hayat, ed.), pp. 237–296. New York: Van Nostrand Reinhold Comp. 1973
Weibel, E.R., Gnägi, H.R.: Improvements in efficiency of Stereologic methods in electron microscopic cytology. In: Electron microscopy, Vol. I (D.S. Bocciarelli, ed.), p. 601. IV. European Conference, Rome (1968)
Weibel, E.R., Gomez, D.M.: A principle for counting tissue structures on random sections. J. appl. Physiol. 17, 343–365 (1962)
Weibel, E.R., Kistler, G.S., Scherle, W.F.: Practical Stereologic methods for morphometric cytology. J. Cell Biol. 30, 23–38 (1966)
Weibel, E.R., Stäubli, W., Gnägi, H.R., Hess, F.A.: Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, Stereologic methods and normal morphometric data for rat liver. J. Cell Biol. 42, 68–91 (1969)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Petrzilka, G.E., Graf-de Beer, M. & Schroeder, H.E. Stereological model system for free cells and base-line data for human peripheral blood-derived small t-lymphocytes. Cell Tissue Res. 192, 121–142 (1978). https://doi.org/10.1007/BF00231028
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00231028