Microwave Immunohistochemistry: Advances in Temperature Control
The advantages and limitations to employing a fiber-optic temperature probe for feedback control of a microwave oven designed for immunohistochemistry were tested. The probe was designed to be submerged in a drop of immunoreagent on a glass slide, and the measured temperature of the droplet was used to regulate the output power and the time needed for the magnetron to reach and maintain the desired incubation temperature. A three-step streptavidin-biotin-peroxidase microwave method, employing a range of antibodies, particularly anti-insulin on rat pancreas, was used as the model system to illustrate the effect of temperature and duration of incubation in microwave immunostaining. Insight into the factors affecting a droplet temperature under microwave irradiation and the relationship among temperature, duration, concentrations of antibodies and immunoreaction for immunostaining were obtained. The optimal temperature and duration for microwave immunostaining in our model system were 37°C for 3 min at 75% power level followed by a 2-min incubation inside the oven for all three stages of immunostaining. Use of droplet temperature-regulated microwave standardizes conditions used in microwave immunohistochemistry. Other potential applications where uniform heating of small amounts of reagent and control of reagent temperature are important may benefit from this technique.
KeywordsHydrogen Peroxide Microwave Convection Paraffin Xylene
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