ULTRASTRUCTURE OF HUMAN LEUKOCYTES AFTER SIMULTANEOUS FIXATION WITH GLUTARALDEHYDE AND OSMIUM TETROXIDE AND "POSTFIXATION" IN URANYL ACETATE

Human leukocytes in suspension or in monolayer cultures have been processed for electron microscopy by fixation in a freshly made cold mixture of glutaraldehyde and osmium tetroxide and by "postfixation" in uranyl acetate. Simultaneous exposure to glutaraldehyde and osmium tetroxide eliminates many of the shortcomings seen when either of these agents is used alone as the initial fixative. Specimens are processed to the stage of dehydration as single cell suspensions or as very small clumps to assure rapid penetration of fixatives and efficient washing. The technique is rapid and reproducible. Electron micrographs presented in this report illustrate the ultrastructural features of human white cells prepared by this method.

[1]  Z. Cohn,et al.  VESICLE FUSION AND FORMATION AT THE SURFACE OF PINOCYTIC VACUOLES IN MACROPHAGES , 1968, The Journal of cell biology.

[2]  Z. Cohn,et al.  AUTOPHAGIC VACUOLES PRODUCED IN VITRO , 1968, The Journal of cell biology.

[3]  Z. Cohn,et al.  AUTOPHAGIC VACUOLES PRODUCED IN VITRO , 1968, The Journal of cell biology.

[4]  I. Watanabe,et al.  Method for electron microscopic studies of circulating human leukocytes and observations on their fine structure. , 1967, Journal of ultrastructure research.

[5]  S. Spicer,et al.  Fine structural studies on the development of heterophil, eosinophil, and basophil granulocytes in rabbits. , 1967, Laboratory investigation; a journal of technical methods and pathology.

[6]  G. Palade,et al.  THE STRUCTURE OF EOSINOPHIL LEUKOCYTE GRANULES IN RODENTS AND IN MAN , 1966, The Journal of cell biology.

[7]  E. Benditt.  The Cell: Its Organelles and Inclusions , 1966 .

[8]  D. R. Anderson.  Ultrastructure of normal and leukemic leukocytes in human peripheral blood. , 1966, Journal of ultrastructure research.

[9]  M. Farquhar,et al.  ORIGIN OF GRANULES IN POLYMORPHONUCLEAR LEUKOCYTES , 1966, The Journal of cell biology.

[10]  L. Weiss,et al.  TRANSFORMATION OF MONOCYTES IN TISSUE CULTURE INTO MACROPHAGES, EPITHELIOID CELLS, AND MULTINUCLEATED GIANT CELLS , 1966, The Journal of cell biology.

[11]  R. Coggeshall,et al.  A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY , 1965, The Journal of cell biology.

[12]  G. de‐Thé,et al.  CYTOPLASMIC MICROTUBULES IN DIFFERENT ANIMAL CELLS , 1964, The Journal of cell biology.

[13]  J. G. Hirsch,et al.  ELECTRON MICROSCOPE STUDIES ON THE DEGRANULATION OF RABBIT PERITONEAL LEUKOCYTES DURING PHAGOCYTOSIS , 1964, The Journal of experimental medicine.

[14]  Y. Tanaka,et al.  FIBRILLAR STRUCTURES IN THE CELLS OF BLOODFORMING ORGANS. , 1964, Journal of the National Cancer Institute.

[15]  A. Novikoff,et al.  GOLGI APPARATUS AND LYSOSOMES. , 1964, Federation proceedings.

[16]  Y. Rabinowitz,et al.  SEPARATION OF LYMPHOCYTES, POLYMORPHONUCLEAR LEUKOCYTES AND MONOCYTES ON GLASS COLUMNS, INCLUDING TISSUE CULTURE OBSERVATIONS. , 1964, Blood.

[17]  W. Lockwood,et al.  ELECTRON MICROGRAPHIC STUDIES OF PHAGOCYTIC CELLS. I. MORPHOLOGICAL CHANGES OF THE CYTOPLASM AND GRANULES OF RABBIT GRANULOCYTES ASSOCIATED WITH INGESTION OF ROUGH PNEUMOCOCCUS. , 1963, British journal of experimental pathology.

[18]  D. Zucker‐Franklin.  THE ULTRASTRUCTURE OF CELLS IN HUMAN THORACIC DUCT LYMPH. , 1963, Journal of ultrastructure research.

[19]  S. de Petris,et al.  Filamentous structures in the cytoplasm of normal mononuclear phagocytes. , 1962, Journal of ultrastructure research.

[20]  H. Florey,et al.  Leucocyte migration from small blood vessels stimulated with ultraviolet light: an electron-microscope study. , 1961, The Journal of pathology and bacteriology.

[21]  John H. Luft,et al.  IMPROVEMENTS IN EPOXY RESIN EMBEDDING METHODS , 1961, The Journal of biophysical and biochemical cytology.

[22]  G. Ackerman,et al.  Morphological and histochemical studies of the leukemic cells from a patient with atypical myeloblastic leukemia with special reference to intracytoplasmic mucopolysaccharide vacuoles and fibrillar formation. , 1960, Blood.

[23]  J. G. Hirsch,et al.  STUDIES OF PHAGOCYTOSIS OF GROUP A STREPTOCOCCI BY POLYMORPHONUCLEAR LEUCOCYTES IN VITRO , 1960, The Journal of experimental medicine.

[24]  E. H. Mercer.  An electron microscopic study of Amoeba proteus , 1959, Proceedings of the Royal Society of London. Series B - Biological Sciences.

[25]  Edward B. Smith.  Electron Microscopic Atlas of Normal and Leukemic Human Blood , 1958 .

[26]  J. A. Freeman,et al.  The ultrastructure of a fibrillar formation of leukemic human blood. , 1958, Blood.

[27]  B. Trump,et al.  New ultrastructural characteristics of cells fixed in a glutaraldehyde-osmium tetroxide mixture. , 1966, Laboratory investigation; a journal of technical methods and pathology.

[28]  C. Duve,et al.  Functions of lysosomes. , 1966, Annual review of physiology.

[29]  D. Fawcett.  The cell, its organelles and inclusions : an atlas of fine structure , 1966 .

[30]  Trump Bf,et al.  New ultrastructural characteristics of cells fixed in a glutaraldehyde-osmium tetroxide mixture. , 1966 .

[31]  J. G. Hirsch.  Neutrophil and Eosinophil Leucocytes , 1965 .

[32]  J. Thiery,et al.  Electron microscopy of human white blood cells and their stem cells. , 1961, International review of cytology.