Immune Electron Microscopy Using a Two‐layer Method of Ferritin Labelling

Baxandall, Jane; Perlmann, Peter; Afzelius, Björn A.

doi:10.1111/j.1365-2818.1963.tb02084.x

Cited by 7 publications

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References 10 publications

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“…The ferritin-labelling technique is readily applicable to the localization of surface antigens, such as the capsular antigens of micro-organisms (Rifkind et al, 1964) and surface antigens of cell membranes (Baxandall et al, 1963), or to the localization of extracellular materials, such as fibrin (White et al, 1964;Wyllie, 1964;Haust et al, 1965). The most useful applications of the technique are likely to arise in the localization of intracellular antigenic sites and it is particularly unfortunate therefore that intracellular localization presents much greater difficulties.…”

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confidence: 99%

Immuno‐electron microscopy of selected cells in tissue sections*

Chapman

Taylor

1966

Journal of the Royal Microscopical Society

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The ultrastructural localization of intracellular antigenic sites in tissue sections presents a number of dlfficulties, particularly when such sites are present in only a small proportion of the total cell population. The method described in this paper depends on finding a suitable fixative (and suitable fixation conditions) that will preserve a reasonable measure of ultrastructure without destroying all immunological activity. Using a doubly-labelled antibody, conjugated with both fluorescein and ferritin, cells in which an immune reaction has occurred can be identified by their fluorescence in the light microscope and these reacting cells can then be selected for subsequent electron microscopy. The thick (15 p) section containing a selected fluorescent cell is prepared for electron microscopy by cutting thin (500 A) sections in the same plane as the original cutting plane and the sites of the immune reaction in the chosen cell can be identified by the presence of the ferritin marker.This immuno-electron optical method has been applied to the study of the cytoplasmic distribution of rheumatoid factor in human lymph node cells and it is shown that the factor can be localized on intracytoplasmic granules.

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confidence: 99%