Detection of plasma cell immunoglobulins in tissue sections optimally fixed for ultrastructural immunocytochemistry.
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.
D000818:Animals; D005557:Formaldehyde; D005976:Glutaral; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D007136:Immunoglobulins / Q000032:analysis*; D008198:Lymph Nodes / Q000166:cytology; D008722:Methods; D051379:Mice; D008854:Microscopy, Electron; D010950:Plasma Cells / Q000276:immunology* / Q000648:ultrastructure; D011108:Polymers; D012996:Solutions
D007136:Immunoglobulins; D011108:Polymers; D012996:Solutions; D005557:Formaldehyde; D005976:Glutaral; C003043:paraform