Distribution of immunoglobulin-producing cells in immunized mouse spleens

Sei Saitoh, Nobuo Terada, Nobuhiko Ohno, Shinichi Ohno

Research output: Chapter in Book/Report/Conference proceedingChapter

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 (Igκ), 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, Igκ- 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, Igκ- or IgG1-immunopositive cells were mostly located in peritrabeculae. The development of Ig-producing cells was clarified in the specimens prepared with IVCT, which proved to be useful for analyzing the native morphology and distribution of Ig-producing cells.

Original languageEnglish
Title of host publicationIn Vivo Cryotechnique in Biomedical Research and Application for Bioimaging of Living Animal Organs
PublisherSpringer Japan
Pages51-56
Number of pages6
ISBN (Electronic)9784431557234
ISBN (Print)9784431557227
DOIs
Publication statusPublished - 01-01-2015

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Saitoh, S., Terada, N., Ohno, N., & Ohno, S. (2015). Distribution of immunoglobulin-producing cells in immunized mouse spleens. In In Vivo Cryotechnique in Biomedical Research and Application for Bioimaging of Living Animal Organs (pp. 51-56). Springer Japan. https://doi.org/10.1007/978-4-431-55723-4_11