Literature DB >> 7400222

The application of polyethylene glycol (PEG) to electron microscopy.

J J Wolosewick.   

Abstract

The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine-coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis.

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Year:  1980        PMID: 7400222      PMCID: PMC2111482          DOI: 10.1083/jcb.86.2.675

Source DB:  PubMed          Journal:  J Cell Biol        ISSN: 0021-9525            Impact factor:   10.539


  12 in total

1.  A rapid imbedding technic for histologic sections employing a water soluble wax.

Authors:  H BLANK
Journal:  J Invest Dermatol       Date:  1949-02       Impact factor: 8.551

2.  Stereo high-voltage electron microscopy of whole cells of the human diploid line, WI-38.

Authors:  J J Wolosewick; K R Porter
Journal:  Am J Anat       Date:  1976-11

3.  Ultrastructural characterization of the manchette microtubules in the seminiferous epithelium of the mouse.

Authors:  J J Wolosewick; J H Bryan
Journal:  Am J Anat       Date:  1977-10

4.  Observation on the morphological heterogeneity of WI-38 cells.

Authors:  J J Wolosewick; K R Porter
Journal:  Am J Anat       Date:  1977-06

5.  Single-step separation of red blood cells. Granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque.

Authors:  D English; B R Andersen
Journal:  J Immunol Methods       Date:  1974-08       Impact factor: 2.303

6.  Light and electron microscopic localization of antigens in tissues embedded in polyethylene glycol with a peroxidase labeled antibody method.

Authors:  J E Mazurkiewicz; P K Nakane
Journal:  J Histochem Cytochem       Date:  1972-12       Impact factor: 2.479

7.  High voltage electron microscopy of critical-point dried cryosection.

Authors:  E Yamada; H Watanabe
Journal:  J Electron Microsc (Tokyo)       Date:  1977

8.  Three dimensional fine structure of cultured cells: possible implications for subcellular motility.

Authors:  I K Buckley
Journal:  Tissue Cell       Date:  1975       Impact factor: 2.466

9.  The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.

Authors:  E S REYNOLDS
Journal:  J Cell Biol       Date:  1963-04       Impact factor: 10.539

10.  Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality.

Authors:  J J Wolosewick; K R Porter
Journal:  J Cell Biol       Date:  1979-07       Impact factor: 10.539

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  18 in total

1.  Imaging cytoskeleton--mitochondrial membrane attachments by embedment-free electron microscopy of saponin-extracted cells.

Authors:  A Lin; G Krockmalnic; S Penman
Journal:  Proc Natl Acad Sci U S A       Date:  1990-11       Impact factor: 11.205

2.  Localization of the adenovirus early region 1B 55-kilodalton protein during lytic infection: association with nuclear viral inclusions requires the early region 4 34-kilodalton protein.

Authors:  D A Ornelles; T Shenk
Journal:  J Virol       Date:  1991-01       Impact factor: 5.103

3.  Distribution of calmodulin in pea seedlings: Immunocytochemical localization in plumules and root apices.

Authors:  M Dauwalder; S J Roux; L Hardison
Journal:  Planta       Date:  1986-09       Impact factor: 4.116

4.  Immunolocalization in three dimensions: immunogold staining of cytoskeletal and nuclear matrix proteins in resinless electron microscopy sections.

Authors:  J A Nickerson; G Krockmalnic; D C He; S Penman
Journal:  Proc Natl Acad Sci U S A       Date:  1990-03       Impact factor: 11.205

5.  Imaging of plant cell walls by confocal Raman microscopy.

Authors:  Notburga Gierlinger; Tobias Keplinger; Michael Harrington
Journal:  Nat Protoc       Date:  2012-08-23       Impact factor: 13.491

6.  Resinless section electron microscopy of HeLa cell mitotic architecture.

Authors:  B Wagner; G Krochmalnic; S Penman
Journal:  Proc Natl Acad Sci U S A       Date:  1986-12       Impact factor: 11.205

7.  Alterations in nuclear matrix structure after adenovirus infection.

Authors:  Z H Zhai; J A Nickerson; G Krochmalnic; S Penman
Journal:  J Virol       Date:  1987-04       Impact factor: 5.103

8.  The cytoskeletal framework of chick osteoclasts in resin-less sections.

Authors:  T Kato; T Akisaka
Journal:  J Anat       Date:  1994-12       Impact factor: 2.610

9.  Arrangement of pili in colonies of Neisseria gonorrhoeae.

Authors:  W J Todd; G P Wray; P J Hitchcock
Journal:  J Bacteriol       Date:  1984-07       Impact factor: 3.490

10.  Detection of fibrils associated with Rickettsia rickettsii.

Authors:  W J Todd; W Burgdorfer; G P Wray
Journal:  Infect Immun       Date:  1983-09       Impact factor: 3.441

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