Immunospecific labeling of mouse lymphocytes in the scanning electron microscope

✍ Scribed by Carter, David P. ;Wofsy, Leon

• Publisher: Wiley (John Wiley & Sons)
• Year: 1976
• Tongue: English
• Weight: 850 KB
• Volume: 5
• Category: Article
• ISSN: 0091-7419
• DOI: 10.1002/jss.400050205

✦ Synopsis

Abstract

Bone marrow‐derived (B) and thymus‐derived (T) Balb/c mouse lymphocytes were identified in the scanning electron microscope (SEM) by the immunospecific attachment of one of several kinds of large‐molecular‐weight markers distinguishable in SEM. These markers (tobacco mosaic virus, keyhole limpet hemocyanin, bushy stunt virus, and bacteriophage T4) could be modified with hapten groups and linked with anti‐hapten antibody, in an indirect (sandwich) scheme, to hapten‐modified anti‐cell‐surface antibody bound to the cell surface.

Hapten‐modified antibodies to B cell antigens (goat anti‐mouse‐immunoglobulin) or to T cell antigens (rabbit anti‐mouse brain) were employed to identify these two lymphoid cell types in unfractionated spleen, mesenteric lymph node, bone marrow, and thymus cell populations. The topography of B cells was always indistinguishable from that of T cells. No surface features were found to be unique to either cell type. In suspension, the majority of B and T cells had one or no microvilli regardless of the tissue source of the labeled cells. Cells in suspension that had microvilli (usually 10% of the total cell population) were always unlabeled. However, after cell contact with a glass surface, approximately half of both the B and T cell populations had a villous topography.