The Journal of Toxicological Sciences
Online ISSN : 1880-3989
Print ISSN : 0388-1350
Regular Paper
COMBINATION OF FIXATION USING PLP FIXATIVE AND EMBEDDING IN PARAFFIN BY THE AMeX METHOD IS USEFUL FOR HISTOCHEMICAL STUDIES IN ASSESSMENT OF IMMUNOTOXICITY
Masami SUZUKIKiyoka KATSUYAMAKenji ADACHIYumie OGAWAKeigo YOROZUEtsuko FUJIIYasuyuki MISAWATetsuro SUGIMOTO
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Volume 27 (2002) Issue 3 Pages 165-172

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Abstract

To establish a method for processing lymphoid organs suited to morphological, immunohistochemical and enzyme histochemical analyses for assessment of immunotoxicity, we examined a combination of fixation with periodate-lysine-paraformaldehyde (PLP) fixative and embedding in paraffin by the AMeX method (PLP-AMeX method). Spleen and thymus removed from monkeys and rats were fixed in PLP fixative for 6 hours at 4°C. After fixation, specimens were processed and embedded in paraffin by the AMeX method. In hematoxylin and eosin-stained sections, tissue architecture was well preserved. In immunohistochemical staining, markers of T lymphocytes (CD3, CD4, CD8), B lymphocytes (monkey: CD20cy, rat: CD45RA) and macrophage (monkey; CD68, rat: ED-1) were well identified according to their specificities, although the staining intensity of CD8 in the monkey and CD4 in the rat were somewhat weaker in PLP-AMeX-prepared sections than in those frozen. In enzyme histochemical staining, alkaline phosphatase activity was well preserved in neutrophils. In toluidine blue- and Giemsa-stained sections, eosinophil granules and the metachromasia of granules in basophil/mast cells were clearly detectable. These findings suggest that the PLP-AMeX method is a powerful tool for assessment of immunotoxicity.

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© 2002 The Japanese Society of Toxicology
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