Ferritin as a Surface Marker for Immunoscanning Electron Microscopy

A method for the localization of intracellular antigens with a scanning electron microscope using peroxidase-labelling antibodies is described. A search for a hydrogen donor which may be deposited at the sites of antigen by enzymatic action and emit secondary electrons or generate backscatter electrons was made. It was found that when 4-chloro-1-naphthol was used, the peroxidase deposited reaction product which resulted in a strong secondary electron emission at the site of antigen. With this method, the presence of luteinizing hormone in secretion granules and other cytoplasmic structures of gonadotropic cells was demonstrated. The level of detection of intracellular antigens with this method is not as high as that detectable with light microscopical examination of the same specimens, that is, more reaction product at the site of antigen is required to be detectable with scanning electron microscopy than with light microscopy. In spite of the lack of high sensitivity, the intracellular antigens may be localized with the method described.

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Immunocytochemical localization of intracellular antigens with SEM

Nakane

Hartman

1980

Histochem J

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Surface Immunoglobulin of Human Lymphocytes

Ito

Hattori

Ito

et al. 1978

Scandinavian Journal of Haematology

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Using ferritin as surface marker, the localization of the surface immunoglobulin (Ig) was studied on peripheral lymphocytes from normal human individuals and patients with macroglobulinaemia Waldenström by scanning immunoelectron microscopy. Normal IgG‐, IgM‐lymphocytes and pathological IgM‐lymphocytes were then compared with regard to their topographical differences. In all cells examined, IgG‐ and IgM‐conjugated ferritin particles were detected all over the cell surface, but the distribution of the former on the normal IgG‐lymphocytes was slightly more diffuse than that of the latter on the normal and pathological IgM‐lymphocytes. Furthermore, in the pathological IgM‐lymphocytes, the clustered IgM‐conjugated ferritin particles were found in great number on the microvilli. Normal IgG‐lymphocytes were almost always characterized by short rod‐like microvilli standing densely and vertically on the cell surface. Some of normal IgM‐lymphocytes had a similar appearance to those of normal IgG‐lymphocytes (type A) but others (type B) had tilted rod‐like microvilli or wide plate‐like processes on their surface. As for IgM‐lymphocytes of macroglobulinaemia, most lymphocytes had tilted rod‐like microvilli and wide plate‐like processes similar to type B, whereas a minor population of the pathological lymphocytes carried long, thin rod‐like microvilli standing vertically on the surface.

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Ferritin as a Surface Marker for Immunoscanning Electron Microscopy

Cited by 2 publications

References 15 publications

Immunocytochemical localization of intracellular antigens with SEM

Immunocytochemical localization of intracellular antigens with SEM

Surface Immunoglobulin of Human Lymphocytes

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