Brief Bio: My name is Tetsuro Inamoto and I’m a veterinarian. I work for Kyoto prefectural office, Japan. While at work, I have been a guest researcher at Kyoto Sangyo University since 2017. I’m interested in the infectious diseases of honeybees and pathological analyses. I’m good at observing the cells microscopically. In my free time I enjoy and learn Japanese tea ceremony.

New method of injecting solution into the abdomen to overcome fixation delay of the midgut on making the histological specimens of honeybee worker

Few histological studies have been performed to evaluate infectious diseases in honeybees, as their water-resistant exoskeletons make the preparation of whole-body specimens difficult by blocking the permeation of fixative and substitution solutions. Additionally, it remains difficult to fix samples when specimens are collected far from the laboratory, as delays in fixation lead to autolysis of abdominal organs. In this study, we propose an intraperitoneal fixative injection method for Apis mellifera to overcome these problems, particularly our method is to inject 30uL fixative solution (3:2 solution of 80% ethanol and Bouin’s fixative) into the abdomen on the day of sampling. This method prevents fixation delay and provides honeybees’ tissue specimens in good condition for histopathological examinations. The results of the injection were favorable by the microscopic observations of the midgut because microvilli were stained well and cellular components corresponding to the terminal web and tight junctions around cell boundaries were clearly distinguished. Besides, this method doesn’t need a temperature control since the storage temperature after fixative injection didn’t affect the results. We concluded this is a simple and effective method. This is the content which I have recently studied in Kyoto Sangyo University. This study is published in Applied Entomology and Zoology (2020) 55: 351-354, introducing as a new method for preparing tissue specimens for histopathological examinations.