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By S. J. Holt and R. Marian Hicks


Discrete localization of stain in pericanalicular granules was found in 10 µ frozen sections of formol-phosphate-sucrose-fixed liver stained by the Gomori acid phosphatase technique and examined in the light microscope. The staining patterns, before and after treatment with Triton X-100 and lecithinase, were identical with those previously reported for formol-calcium-fixed material treated in the same way, and it can be assumed that the stained granules are identical with "lysosomes." Examination in the light microscope of the staining patterns and lead penetration in fixed blocks and slices of various dimensions showed nuclear staining and other artefacts to be present, produced by the different rates of penetration of the various components of the staining medium into the tissue. A uniform pericanalicular staining pattern could be obtained, however, with slices not more than 50 µ thick, into which the staining medium could penetrate rapidly from both faces. The staining pattern produced in 50 µ slices was the same both at pH 5.0 and pH 6.2, and was not altered by subsequent embedding of the stained material in butyl methacrylate. Electron microscopy showed the fine structure of fixed 50 µ frozen slices to be well preserved, but it deteriorated badly when they were incubated in the normal Gomori medium at pH 5.0 before postfixing in osmium tetroxide. After incubation in the Gomori medium at pH 6.2, the detailed morphology was substantially maintained. In both cases lead phosphate, the reaction product, was found in the pericanalicular regions of the cell, but only in the vacuolated dense bodies and never in the microbodies. Not every vacuolated dense body contained lead, and stained and unstained bodies were sometimes seen adjacent to each other. This heterogeneous distribution of stain within a morphologically homogeneous group of particles is consistent with de Duve's suggestion (9) that there is a heterogeneous distribution of enzymes within the lysosome population. It is concluded from these investigations that the vacuolated dense bodies seen in the electron microscope are the morphological counterparts of the "lysosomes" defined biochemically by de Duve

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    1. (1959). A diazo coupling method for the electron microscopic localization of cholinesterase,
    2. A suggestion as to the nature of the lysosome granules,
    3. (1959). Activity of the isolated membranes of the Golgi system,
    4. (1958). Adenosine-triphosphatase and 5-nucleotidase activities in the plasma membrane of liver cells as revealed by electron
    5. (1959). An electron microscope study of the early effects of 3'-MeDAB on rat liver cells, Cancer Research,
    6. (1958). Application of histochemistry to electron microscopy,
    7. (1960). Biochemical and staining reactions of cytoplasmic constituents,
    8. (1960). Degranulation of polymorphonuclear leucocytes following phagoeytosis of microorganisms,
    9. (1951). Effect of formalin fixation on the activity of five enzymes of rat liver,
    10. (1957). Effects of varying the vehicle for OsO4 in tissue fixation,
    11. Electron microscopic observations on developing chick embryo liver. The Golgi complex and its possible role in the formation of glycogen,
    12. (1956). Electron microscopy of lysosome-rich fractions from rat
    13. (1959). Factors governing the validity of staining methods for enzymes, and their bearing upon the Gomori acid phosphatase technique, Exp. Cell Research,
    14. (1955). Histochemical reactions for electron microscopy : Acid phosphatase, Exp. Cell Research,
    15. (1950). Histochemical studies on tissue enzymes. V. A difficulty in enzyme localization in the acid range due to selective affinity of certain tissues for lead; its dependence on pH,
    16. (1956). Liver microsomes. An integrated morphological and biochemical
    17. (1959). Lysosomes, a new group of cytoplasmic particles, in Subcellular Particles,
    18. Microbodies" and the problem of mitochondrial regeneration in liver ceils,
    19. (1952). Microscopic tlistochemistry; Principles and Practice,
    20. (1939). Microtechnical demonstration of phosphatase in tissue sections,
    21. (1934). Phosphatase activity of animal tissues,
    22. (1960). Preservation of integrity of rat tissues for cytochemical staining purposes,
    23. (1960). quoted by de Duve in report on Colloquium on Intracellular Localization of Enzymes, Louvain,
    24. (1956). Some problems in localizing enzymes at the electron microscope
    25. (1958). Staining of tissue sections fo~ electron microscopy with heavy metals. II. Application of solutions containing lead and
    26. (1958). Studies in enzyme cytochemistry. I. Principles of cytoS.
    27. (1961). Studies on formalin fixation for electron microscopy and cytochemical staining purposes,
    28. (1957). The acid phosphatases of rat liver,
    29. (1959). The demonstration with the electron microscope of the end-products of histochemical reactions in relation to the fine structure of cells,
    30. (1952). The fixation of tissues for electron microscopy,
    31. (1946). The histochemical recognition of lipine,
    32. (1960). The liver cell. Some new approaches to its study,
    33. (1959). The localization of adenosine triphosphatase activity in rat kidney: Electron microscopic examination of reaction product in formol-calcium-fixed frozen sections,
    34. (1936). The structure of animal fibres in relation to acid dyeing,
    35. Theoretical and Applied,
    36. (1955). Tissue fractionation studies. 6. Intracellular distribution pattelns of enzymes in rat-liver tissue,
    37. (1959). Tissue fractionation studies. 9. Release of bound hydrolases,
    38. (1961). Use of veronal buffers in formalin fixatives, Nature,

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