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1994-09-19
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Document 0466
DOCN M9490466
TI A morphological and immunolabelling study of freeze-substituted human
and simian immunodeficiency viruses.
DT 9411
AU Grief C; Nermut MV; Hockley DJ; Electron Microscopy and Photography
Section, National Institute; for Biological Standards and Control,
Potters Bar, Herts, U.K.
SO Micron. 1994;25(2):119-28. Unique Identifier : AIDSLINE MED/94332554
AB Rapid freezing, freeze substitution and low temperature embedding were
used to obtain resin-embedded specimens of HIV and SIV for morphological
and immunolabelling studies, with particular emphasis on the 'lateral
bodies' and p6 protein. HIV- or SIV-infected cells were fixed in 3%
paraformaldehyde and cryoprotected with 0.5 M sucrose. Cells were
applied to pieces of Whatman No 1 filter paper and impact-frozen onto a
liquid nitrogen cooled copper block. Specimens were freeze-substituted
at -90 degrees C using one of three different media: (a) absolute
methanol, (b) methanol containing 0.5% uranyl acetate, and (c) methanol
containing glutaraldehyde, osmium tetroxide and uranyl acetate.
Specimens substituted in methanol and uranyl acetate showed both good
structural preservation and retention of antigenicity. We found that the
use of filter paper for supporting the specimen was an important factor
in obtaining good freezing rates and was more practical than freezing
mixtures of cells and gelatin. When compared with specimens prepared by
conventional fixation and embedding, freeze-substituted virus particles
showed a greater uniformity of shape and size and were more dense in
appearance. Distinct 'lateral bodies' were not observed in
freeze-substituted viruses. The viral protein p6 was widely distributed
in the centre of mature virus particles.
DE Animal Antibodies, Monoclonal Comparative Study Freeze
Substitution/*METHODS Human HIV/CHEMISTRY/*ULTRASTRUCTURE
Immunohistochemistry Microscopy, Electron
SIV/CHEMISTRY/*ULTRASTRUCTURE Tissue Embedding/METHODS Viral
Proteins/ANALYSIS JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).