Microwave fixation: understanding the variables to achieve rapid reproducible results.
The use of microwave irradiation for rapid chemical fixation of tissues in electron microscopy is a subject of current interest. The effects of water load size and location, sample placement in the oven cavity (hot or cold spots), and time on tissue preservation were examined. The use of a microwave container (4 dram vial) encased in 60 ml of ice in a 100 ml polyethylene beaker and a 0% power setting between two 100% power settings (time interval) provided reliable control of temperature during microwave irradiation. High brightness neon lights provided a quick and easy method to identify and map hot and cold spots within the oven cavity. Using microwave irradiation for rapid glutaraldehyde and osmium tetroxide fixation of tissues (Pacific yew needle and mouse kidney and liver) for electron microscopy yielded preservation equal or better than routine immersion fixation when a time interval, a cold spot (as the sample location), and an ice-encased vial were used during microwave fixation. These adaptations provided reliable control of fixation conditions in an 800 watt laboratory microwave oven.
Giberson
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Ted Pella, Inc., Redding, California 96049, USA.
Demaree
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RS
D000818:Animals; D002736:Chloroplasts / Q000648:ultrastructure; D004574:Electromagnetic Fields; D051379:Mice; D008854:Microscopy, Electron; D008872:Microwaves; D015203:Reproducibility of Results; D016707:Tissue Fixation / Q000379:methods*