Detection of plasma cell immunoglobulins in tissue sections optimally fixed for ultrastructural immunocytochemistry

• Published in: The journal of histochemistry and cytochemistry Vol. 35; no. 2; pp. 189 - 196
• Main Authors: Gonatas, NK, Gonatas, JO, Stieber, A, Ternynck, T, Avrameas, S
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA Histochemical Soc 01.02.1987; SAGE Publications; Histochemical Society
• Subjects: Analysis of the immune response. Humoral and cellular immunity; Animals; Biological and medical sciences; Formaldehyde; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Glutaral; Histocytochemistry; Immunobiology; Immunoenzyme Techniques; Immunoglobulins - analysis; Lymph Nodes - cytology; Methods; Mice; Microscopy, Electron; Organs and cells involved in the immune response; Plasma Cells - immunology; Plasma Cells - ultrastructure; Polymers; Solutions; Immunoglobulins; Lymph node; Immunocytochemistry; Rodentia; Plasmocyte; Enzyme immunoassay; Biotine; Immunoperoxidase technique; Golgi apparatus; Histological fixation; Vertebrata; Mammalia; Transmission electron microscopy; Lymphoid organ; Mouse; Animal
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1177/35.2.3098833

We describe the ultrastructural localization of plasma cell immunoglobulins in vibratome sections of popliteal lymph nodes. Fixation with glutaraldehyde-paraformaldehyde gave better tissue and antigen preservation than paraformaldehyde or periodic acid lysine-paraformaldehyde; biotinylated Fab fragments of sheep anti-mouse IgG-streptavidin-biotinylated horseradish peroxidase (HRP) or Fab-HRP conjugates gave similar results. With both immunoreagents, excellent tissue preservation and antigen detection was observed in the first layer of cells sectioned with the vibratome. Conjugates of anti-mouse IgG with HRP did not show any staining. Peroxidase stain was observed in the nuclear envelope, cisternae of the rough endoplasmic reticulum, and the Golgi apparatus complex. In the Golgi apparatus, staining was seen consistently in cisternae of the cis face and in adjacent vesicles; the trans cisternae showed weak or no stain, and adjacent vesicles, "coated" vesicles, and granules were not stained. This study shows that high quality of tissue preservation and antigen detection, by both light and ultrastructural immunocytochemistry, is feasible in tissue fixed with glutaraldehyde-paraformaldehyde followed by vibratome sectioning and immunostaining with Fab-biotin-streptavidin-biotin-HRP, or Fab-HRP.