Microwave staining of enteric neurons using cuprolinic blue (quinolinic phthalocyanin) combined with enzyme histochemistry and peroxidase immunohistochemistry

Methods that visualize subsets as well as the entire enteric neuron populat ion are not readily available or have proved to be unreliable. Therefore, w e attempted to combine NADPH-d histochemistry, AChE histochemistry, and CGR P immunohistochemistry, techniques that mark subsets of enteric neurons, wi th a technique that appeared to visualize the entire enteric neuron populat ion, the cuprolinic blue staining method. To guarantee representative stain ing results, the individual staining methods were modified by using microwa ves. In addition, this preserved the characteristics of each of the individ ual techniques. The distribution of NADPH-d, AChE, and CGRP corresponded we ll with previous morphological and physiological reports. Consequently, the different combinations gave rise to rapid, useful, and ready-to-use double labeling techniques. Their main advantage is that they simultaneously Visu alize the total population as well as subsets of enteric neurons.