Application of microwave fixation techniques in pathology to neuroscience studies: a review

The introduction of microwave energy into the scientist's repertoire of fixation modalities offers for the first time in relatively large specimens the potential for 'instantaneous' preservation of cellular structure for light and electron microscopy with minimal alteration of cellular biochemistry and antigenicity. Because of the rapid evolution of this new technology, we provide a classification system of newly generated microwave methods as applied to specimen preservation for microscopic analysis. With emphasis on neuronal tissue, we review qualitative and quantitative microscopy data of specimens fixed by two microwave methods in common use: (1) microwave stabilization and (2) fast and ultrafast, primary microwave-chemical fixation. In addition, we provide a table of neuropeptides or proteins in neuronal tissues that are preserved by various microwave fixation methods for histochemistry, immunohistochemistry, and immuno-electron microscopy studies. Commercial microwave ovens have limitations which can result in irreproducible fixation results. Therefore, we present a calibration protocol that is used to identify the best locations for fixation within large cavity (i.e., household) microwave ovens. We also provide a standardization protocol to improve the reproducibility of microwave fixation in calibrated, large-cavity microwave ovens.