摘要:

AbstractThe properties of the inorganic dye ruthenium red are presented with emphasis upon its use for electron microscopy of cells and tissues. Although commercial ruthenium red often can be used directly, it always contains various impurities and by‐products. One of these, termed ruthenium violet, can be isolated and is useful by itself. Absorption spectra of the ruthenium dyes and common impurities are given so that an assay is possible for any sample. Convenient fixative recipes containing ruthenium red or violet are provided together with constraints necessary for a reliable reaction to label extracellular acidic mucosubstances. Perfusion was not successful. The specificity of the ruthenium red reaction was evaluated by spot testing with 57 substances, and by titration with chemically defined pectins. The results indicate that ruthenium red, as a hexavalent cation, precipitates a large variety of polyanions by ionic interaction, and that its classical reaction with pectin is typical rather than specific. New data are presented regarding its reaction with phospholipids. For electron microscopy, a further reaction with OsO4amplifies the feeble electron density, which is the counterpart of its intense optical labeling, to a practical level resulting in strong contrast. An hypothesis is presented for the mechanism underlying this intensificatio

ISSN:0003-276X    

DOI:10.1002/ar.1091710302

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1971

数据来源: WILEY

摘要:

AbstractThe inorganic dye, ruthenium red, stains extracellular materials in animal tissues which probably are acidic mucopolysaccharides. It complements other techniques, its advantages being fine grain, high resolution and good contrast. Localization is shown in mouse and rat muscle, heart, lung and intestine, frog cartilage and cells scraped from oral epithelium of human beings. Attention is paid to collagen bundles, the cell/collagen interface and particularly the myotendinal junction, cartilage matrix and agar gel, desmosomes, intestinal microvilli, erythrocytes and vascular endothelium, nerve fibers and the T‐system of striated muscle. Although ruthenium red generally is excluded by plasma membranes, it penetrates giving intracellular density, if the membrane is broken. Even when the cell membrane is intact, exceptions occur with selective staining of the T‐tubules or the sarcoplasmic sacs depending upon the state of contraction of the muscle cell, and with intracellular staining of certain nuclei and epithelial cells. Ruthenium red stains intracellular lipid droplets revealing lamellae, and stains myelin forms grown from crude egg lecithin but cannot penetrate deeply. It is localized in extracellular materials which have an important mechanical function. Its exclusion by cell membranes permits tracing tortuous cellular invaginations and those exceptions to its exclusion invite a comparison of the localization of the dye with the function of the c

ISSN:0003-276X    

DOI:10.1002/ar.1091710303

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1971

数据来源: WILEY

摘要:

AbstractRuthenium violet, closely related to ruthenium red, supplements the ultrastructural knowledge of the plasma membrane complex. Amoebae throughout were handled individually with braking pipettes and were exposed to ruthenium violet alive, during fixation with acrolein and OSO4, or during dehydration. Ruthenium violet was less toxic than ruthenium red but still killed the amoebae. Conventional methods reveal a filamentous layer 2000 Å thick, an amorphous layer 150 Å thick, and a typical trilaminar plasma membrane (48 Å center‐to‐center). Ruthenium violet binds to the plasma membrane, and to the extraneous coats revealing globules in the filamentous layer. The diameter of the globules decreased according to the stage of processing at which the amoebae first encountered ruthenium violet; they were 1200 Å in diameter when amoebae were alive, 600 Å in acrolein and 300 Å in dehydration. The appearance of the filamentous layer varied when ruthenium violet was replaced by very pure ruthenium red or red containing ruthenium brown (typical of commercial ruthenium red). The globules could be demonstrated without using ruthenium dyes when amoebae were treated after fixation with uranyl acetate or phosphotungstic acid. The relationship of extraneous coats of amoebae is compared with the coats and laminae of animal tis

ISSN:0003-276X    

DOI:10.1002/ar.1091710304

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1971

数据来源: WILEY

ISSN:0003-276X    

DOI:10.1002/ar.1091710301

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1971

数据来源: WILEY