HISTAMINE IMMUNOCYTOCHEMISTRY ON ROUTINE PARAFFIN-EMBEDDED SECTIONS IN THE RAT STOMACH

• Published in: Toxicology mechanisms and methods 1998-01, Vol.8 (2), p.151-163
• Main Author: Matsuzawa, Takao Shishido Toshiaki
• Format: Article
• Language: English
• Subjects: Fixation Histamine Storing Cell Immersion Immunocytochemistry Microwave Irradiation Paraffin Embedding Section
• ISSN: 1537-6516; 1051-7235; 1537-6524
• DOI: 10.1080/105172398242970

A technique has been adapted from methods that maintain the same staining quality as that produced by perfusion fixation and immunocytochemistry on frozen sections of the stomach, and that can be easily incorporated into the routine procedures for paraffin embedded sections in toxicity studies in the rat. The technique is as follows: The rat is killed by exsanguination under ether anesthesia and the stomach is pinned on a corkboard. The stomach is then immediately immersed in 4% 1-ethyl-3(3-dimethyle aminoprophyl)carbodiimide in 0.1 M phosphate-buffered solution, pH 7.4, at 4 C for 2 h and then in 4% paraformaldehyde in the same buffer solution for 2 h. The tissue is irradiated by microwaves twice at 250 W for 20 s each time. The tissue is then embedded in paraffin wax at 58 to 60 C. This technique has proved suitable for staining histamine-containing cells in the stomach and can be incorporated as part of the routine necropsy on routine large toxicity studies. The process has improved both the identification of histamine-containing cells and the storage quality of the fixed tissue. Microwave irradiation and storage of the sample at 4 C in 80% EtOH for 12 weeks did not affect the staining affinity. These methods have reduced the workload by 30 to40%compared to the procedureof perfusion fixation and frozen section.