Guide to Clinical and Diagnostic Virology

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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A combined pseudoreplica-immunocytochemical technique for research and diagnostic virology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162065 ◽

1990 ◽

Vol 48 (3) ◽

pp. 900-901

Author(s):

Blayne Fritz ◽

Stanley J. Naides ◽

Kenneth Moore

Keyword(s):

Phosphotungstic Acid ◽

Immunogold Labelling ◽

Uranyl Acetate ◽

Distilled Water ◽

Clinical Specimens ◽

Tissue Sections ◽

Specimen Retrieval ◽

Transmission Electron ◽

Viral Particles ◽

Diagnostic Virology

The pseudoreplica method of staining viral particles for visualization by transmission electron microscopy is a very popular technique. The ability to concentrate clinical specimens while semi-embedding viral particles makes it especially well suited for morphologic and diagnostic virology. Immunolabelling viral particles with colloidal gold is a technique frequently employed by both research and diagnostic virologists. We have characterized a procedure which provides the advantage of both by modifying and combining pseudoreplica staining and immunogold labelling.Modification of specimen retrieval and delay of staining allows us to utilize pseudoreplica processed specimens within our standard immunogold labelling protocol. In brief, we absorbed samples onto 2% agarose, added.25% Formvar and wicked dry. We then floated the Formvar-virus film onto double distilled water, added grids and retrieved with parafilm. The Formvar-virus specimens were then treated as thin tissue sections within our standard two stage immunolabelling protocol. Following completion of immunogold labelling; each grid was negatively stained with phosphotungstic acid or uranyl acetate contrast stains.

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