Tissue Culture Conservation of Woody Species

The morphologic pathologist may require information on the ultrastructure of a non-specific lesion seen under the light microscope before he can make a specific determination. Such lesions, when caused by infectious disease agents, may be sparsely distributed in any organ system. Tissue culture systems, too, may only have widely dispersed foci suitable for ultrastructural study. In these situations, when only a few, small foci in large tissue areas are useful for electron microscopy, it is advantageous to employ a methodology which rapidly selects a single tissue focus that is expected to yield beneficial ultrastructural data from amongst the surrounding tissue. This is in essence what "LIFTING" accomplishes. We have developed LIFTING to a high degree of accuracy and repeatability utilizing the Microlift (Fig 1), and have successfully applied it to tissue culture monolayers, histologic paraffin sections, and tissue blocks with large surface areas that had been initially fixed for either light or electron microscopy.

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Recent Studies on a Murine Leukemia Virus Grown in Tissue Culture

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060544 ◽

1967 ◽

Vol 25 ◽

pp. 106-107

Author(s):

L. Z. de Tkaczevski ◽

E. de Harven ◽

C. Friend

Keyword(s):

Tissue Culture ◽

Solid Tumors ◽

Murine Leukemia Virus ◽

Leukemia Virus ◽

Supernatant Fluid ◽

Virus Particles ◽

Friend Leukemia ◽

Type C ◽

Leukemia Viruses ◽

Murine Leukemia

Despite extensive studies, the correlation between the morphology and pathogenicity of murine leukemia viruses (MLV) has not yet been clarified. The virus particles found in the plasma of leukemic mice belong to 2 distinct groups, 1 or 2% of them being enveloped A particles and the vast majority being of type C. It is generally believed that these 2 types of particles represent different phases in the development of the same virus. Particles of type A have been thought to be an earlier form of type C particles. One of the tissue culture lines established from Friend leukemia solid tumors has provided the material for the present study. The supernatant fluid of the line designated C-1A contains an almost pure population of A particles as illustrated in Figure 1. The ratio is, therefore, the reverse of what is unvariably observed in the plasma of leukemic mice where C particles predominate.

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Cytoplasmic Tubules in Cells Infected with Laryngotracheitis Virus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100063792 ◽

1969 ◽

Vol 27 ◽

pp. 376-377

Author(s):

A. M. Watrach

Keyword(s):

Tissue Culture ◽

Small Proportion ◽

Chicken Embryo ◽

Culture Cells ◽

Tissue Culture Cells ◽

Morphologic Characteristics ◽

Infectious Laryngotracheitis ◽

Laryngotracheitis Virus ◽

In Cells

During a study of the development of infectious laryngotracheitis (LT) virus in tissue culture cells, unusual tubular formations were found in the cytoplasm of a small proportion of the affected cells. It is the purpose of this report to describe the morphologic characteristics of the tubules and to discuss their possible association with the development of virus.The source and maintenance of the strain of LT virus have been described. Prior to this study, the virus was passed several times in chicken embryo kidney (CEK) tissue culture cells.

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Uranyl Acetate and Rotary Shadowing to Increase the Contrast of Small Aggregated Virus Particles

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100064037 ◽

1969 ◽

Vol 27 ◽

pp. 424-425

Author(s):

Lee F. Ellis ◽

Richard M. Van Frank ◽

Walter J. Kleinschmidt

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Density Gradient ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Uranyl Acetate ◽

Interferon Inducer ◽

Virus Particles ◽

Staining Methods ◽

Rotary Shadowing

The extract from Penicillum stoliniferum, known as statolon, has been purified by density gradient centrifugation. These centrifuge fractions contained virus particles that are an interferon inducer in mice or in tissue culture. Highly purified preparations of these particles are difficult to enumerate by electron microscopy because of aggregation. Therefore a study of staining methods was undertaken.

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A Method for Electron Microscopic Study of Tissue Culture Cell Monolayers Grown in Tubes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060672 ◽

1967 ◽

Vol 25 ◽

pp. 132-133

Author(s):

R. Stephens ◽

G. Schidlovsky ◽

S. Kuzmic ◽

P. Gaudreau

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Light Microscopy ◽

Cell Sheet ◽

Culture Cell ◽

Tissue Culture Cell ◽

Electron Microscopic ◽

Cell Sheets ◽

Morphological Alterations ◽

Culture Cells

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.

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Electron microscopy in diagnostic virology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100110702 ◽

1984 ◽

Vol 42 ◽

pp. 70-73

Author(s):

S. E. Miller

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Molecular Identification ◽

Point Of View ◽

Negative Stain ◽

Viral Diagnosis ◽

Electron Microscopist ◽

Diagnostic Virology

The techniques for detecting viruses are many and varied including FAT, ELISA, SPIRA, RPHA, SRH, TIA, ID, IEOP, GC (1); CF, CIE (2); Tzanck (3); EM, IEM (4); and molecular identification (5). This paper will deal with viral diagnosis by electron microscopy and will be organized from the point of view of the electron microscopist who is asked to look for an unknown agent--a consideration of the specimen and possible agents rather than from a virologist's view of comparing all the different viruses. The first step is to ascertain the specimen source and select the method of preparation, e. g. negative stain or embedment, and whether the sample should be precleared by centrifugation, concentrated, or inoculated into tissue culture. Also, knowing the type of specimen and patient symptoms will lend suggestions of possible agents and eliminate some viruses, e. g. Rotavirus will not be seen in brain, nor Rabies in stool, but preconceived notions should not prejudice the observer into missing an unlikely pathogen.

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Ultrastructure of two neoplasms in culture compared with original biopsies

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100110623 ◽

1984 ◽

Vol 42 ◽

pp. 50-51

Author(s):

Joseph M. Harb ◽

James T. Casper ◽

Vlcki Piaskowski

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Biopsy Specimen ◽

Cultured Cells ◽

Light And Electron Microscopy ◽

Human Tumor ◽

Soft Agar ◽

Human Tumor Cells ◽

Morphological Distinction ◽

Tumor Types

The application of tissue culture and the newer methodologies of direct cloning and colony formation of human tumor cells in soft agar hold promise as valuable modalities for a variety of diagnostic studies, which include morphological distinction between tumor types by electron microscopy (EM). We present here two cases in which cells in culture expressed distinct morphological features not apparent in the original biopsy specimen. Evaluation of the original biopsies by light and electron microscopy indicated both neoplasms to be undifferentiated sarcomas. Colonies of cells propagated in soft agar displayed features of rhabdomyoblasts in one case, and cultured cells of the second biopsy expressed features of Ewing's sarcoma.

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