A Simple Rapid Method for Staining Epoxy-Embedded Specimens for Light Microscopy with the Polychromatic Stain Paragon-1301

Spurlock, Ben O.; Skinner, Margaret S.; Kattine, Anthony A.

doi:10.1093/ajcp/46.2.252

Cited by 110 publications

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“…Since permanganate-fixed tissue swelled during polymerization, all permanganatefixed tissue was embedded in Maraglas according to the method of Spurlock et al (26).…”

Section: Methodsmentioning

confidence: 99%

THE ULTRASTRUCTURE OF COTYLEDONARY TISSUE FROM GOSSYPIUM HIRSUTUM L. SEEDS

Yatsu

1965

The Journal of Cell Biology

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Quiescent cottonseeds stored in a dry, anaerobic situation for over a year have been shown to contain cells whose contents are ultrastructurally similar to those of normal, fully hydrated plant cells. Plastids, mitochondria, and nuclei of the cells of cotyledon tissue in dry seeds possess normal-looking double membranes even under conditions of extreme desiccation. Previous reports have indicated on the basis of light microscopic work, that the cells of certain dry seeds do not possess nuclear membranes or mitochondria. The cells of the dry cottonseed do contain these structures, however. Dictyosomes have not been observed in the spongy parenchymal cells of the cotyledon tissue; it is suggested that they are concerned with translocation and/or utilization of material. The storage materials in the cells, protein and oil, are contained in vacuolar areas enclosed by a single membrane.Seeds offer a convenient base line in ontogenetic studies of the plant. Many ultrastructural studies have been directed to the terminal stages of seed development (7,8,12,17,27) or to the initial stages of seed germination (4,10,18,22,28), but relatively few studies have been conducted on the dry seed per se. Yet an understanding of the final changes on maturation and the first events of germination requires information on the mature, quiescent seed as a point of reference.Studies of biochemical changes in seeds which have initiated germination processes (1, 9, 31) show a dramatic rise in activity of many enzymes within a very short time. Wasserman (29), working with light microscopy, was unable to see mitochondria in cells of the dry seed. However, within 12 to 24 hours after the seeds had imbibed water, mitochondria in cells became visible, presumably arising de novo. Thus, it is obvious that changes occur quite rapidly in seeds after imbibition of water.Unfortunately, most electron microscope studies have been conducted on seeds which were allowed to imbibe water for various periods prior to fixation (16,18,25,28). Since rapid changes do occur in seeds upon imbibition of water, any technique involving imbibition of water, even for short periods, may not reflect their original condition. The paucity of studies on dry seeds is probably due to the extreme difficulty of working with them. This study, therefore, was undertaken to determine the structure of cells in dry, quiescent seeds fixed directly without prior hydration. MATERIALS AND METHODSA master sample of cottonseed (Gossypium hirsutum L., var. Acala 4-42-77, Glandless) harvested in Fall, 1962 was stored in a vacuum desiccator with a desiccant at 3°C and subsamples were removed as needed. Sections of tissue approximately 0.3 mm in diameter were cut carefully from the seed and fixed immediately in two different fixatives, osmium tetroxide and permanganate salts. OSMIUM :Since earlier work indicated that various solvents deranged the normal structure of the dry

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“…Since permanganate-fixed tissue swelled during polymerization, all permanganatefixed tissue was embedded in Maraglas according to the method of Spurlock et al (26).…”

Section: Methodsmentioning

confidence: 99%

THE ULTRASTRUCTURE OF COTYLEDONARY TISSUE FROM GOSSYPIUM HIRSUTUM L. SEEDS

Yatsu

1965

The Journal of Cell Biology

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“…Monotcrpenes and scsq uiterpcne~ are characteristie components of this oil, being the major constituents cincole ( eucaliptol), camphor awl a-pinene. (Spurlock et al, 1966). Frozen sections for lipid hi,;tochcmi~try were stained bv Swlan blaek B and Sudan Ill.…”

Section: * ** Introductionmentioning

confidence: 99%

Secretory Trichomes fromArtemisia Crithmifolia: some ultrastructural aspects

Ascensão¹,

Pais

1982

Bulletin de la Société Botanique de France. Actualités Botaniqu

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.Summary.-The secretory trichomes from Artcmi.fia crithmifolia present a I.Jiscriate multicellular structure. They originale from a single epidrrmal cell, belonging to the leaf orto the ~tem primordia. Histochcmical tests reveal the lipophilic character of the secreted matcrial corresponding probably to terpenoids.Young trichome cells possess the normal features of a secretory cel!, namely a dense cylo· plasm conlaining numerous mitochondria and plastids and few vacuoles. A large nucleus with one or more prominenl nucleoli is also present.Modifications of plastids morphology and increase of ER profiles must be rclated to sccre· tion. Osmiophilic droplets are seen in close relation with plastids and ER. Electron dense matcrial has also been observed in small vacuoles, corresponding presumably to the storage of secretion.Secretory products are extruded and temporarily accumulatcd in the subcuticular space. Older trichomes undergo a lytic process followed by trichomes death.Résumé.-Les trichomes sécréteurs d'Artemisin crithmifolia sont formés par plmieurs cd· Iules à disposition bisériée. Ils proviennent d'une cellule épidermique appartenant soit à la feuille soit à la tige. Des tests histochimiques révèlent la nature lipophilique de la sécrétion. Il s'agit proba· blement de terpènes.Les cellules des jeunes trichomes présentent l'aspect caractéristique des cellules sécrétrice:.:, notamment un noyau très volumineux à un ou plusieurs nucléoles et un cytoplasme riche en ribo·

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“…Sections 2 µm thick were stained with methylene blue and basic fuchsine (BENNETT et al 1976) or toluidine blue (SPURLOCK et al 1966). Measurements on the serial sections stained with trichrome methods were made by means of a calibrated eyepiece micrometer.…”

Section: Methodsmentioning

confidence: 99%

Histological and histochemical characterization of secretory cells of the male copulatory organs of Girardia anderlani (Platyhelminthes: Tricladida: Paludicola)

Vara

Leal-Zanchet

2008

Rev. Bras. Zool.

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Glands of the reproductive system are important for taxonomical identification of flatworms. We studied the histology and histochemical characteristics of the glands of the male copulatory apparatus in Girardia anderlani (KAWAKATSU & HAUSER, 1983). Specimens were fixed in reproductive state, i. e. during and following copulation at four, eight, 12 and 16 hours intervals. Secretory cells were distinguished on the basis of secretion morphology and their staining properties, using trichrome methods and histochemical reactions. Twelve secretory cell types and five main types of secretions were identified in the male copulatory apparatus: glycoproteic with and without tryptophan; glycosaminoglycidic; neutral mucopolysaccharidic; and proteic. Compared to other Girardia species, more diverse types of secretory cells comprise the glands of the male reproductive system. Histophysiological comparative studies of species of Girardia, in a reproductive state, are necessary to characterize the various regions of the copulatory apparatus as well as to understand the physiology of reproduction.

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A Simple Rapid Method for Staining Epoxy-Embedded Specimens for Light Microscopy with the Polychromatic Stain Paragon-1301

Cited by 110 publications

References 0 publications

THE ULTRASTRUCTURE OF COTYLEDONARY TISSUE FROM GOSSYPIUM HIRSUTUM L. SEEDS

THE ULTRASTRUCTURE OF COTYLEDONARY TISSUE FROM GOSSYPIUM HIRSUTUM L. SEEDS

Secretory Trichomes fromArtemisia Crithmifolia: some ultrastructural aspects

Histological and histochemical characterization of secretory cells of the male copulatory organs of Girardia anderlani (Platyhelminthes: Tricladida: Paludicola)

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