Distribution of immunoglobulin-producing cells in immunized mouse spleens revealed with "in vivo cryotechnique"

Journal of Immunological Methods
S SaitohS Ohno

Abstract

To identify immunoglobulin (Ig)-producing cells with immunohistochemistry, conventional methods of preparation using chemical fixatives have problems such as the artificial diffusion of components and antigen masking. The "diffusion artifact" is caused by the translocation of soluble proteins like Ig from the serum to cytoplasm or vice versa. We have examined the immunolocalization of serum proteins, such as Ig kappa light chain (Igkappa), IgG1 heavy chain (IgG1), and albumin, in immunized mouse spleens after a peritoneal injection of human hemoglobin. Better preservation of morphology and immunoreactivity was obtained with the "in vivo cryotechnique" (IVCT) followed by freeze-substitution, than with conventional preparative methods. Although Ig-producing cells were not clearly detected in red pulp of 2-day-immunized spleens with the conventional methods, Igkappa-immunopositive cells with rich cytoplasm were detected in the red pulp with IVCT, especially in the subcapsular and peritrabecular areas, where IgG1-immunopositive cells were rarely observed. In 7-day-immunized spleens prepared with IVCT, Igkappa- or IgG1-immunopositive cells were mostly located in peritrabeculae. The development of Ig-producing cells was clarified in ...Continue Reading

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Citations

Jul 30, 2010·Journal of Electron Microscopy·Shinichi OhnoYasuhisa Fujii
May 28, 2009·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Nobuo TeradaShinichi Ohno
Jul 25, 2014·Medical Molecular Morphology·Seiichiro HemmiAkihiro Hemmi
May 28, 2020·International Journal of Surgical Pathology·Rie NakataHiroyoshi Ota
Jan 29, 2021·International Journal of Molecular Sciences·Yuta SakamotoSatoshi Shimo

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