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Journal of Histochemistry and Cytochemistry, Vol. 47, 13-22, January 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Microwave Staining of Enteric Neurons Using Cuprolinic Blue (Quinolinic Phthalocyanin) Combined with Enzyme Histochemistry and Peroxidase Immunohistochemistry

Christa J. Van Ginnekena, Marcel J. De Smeta, Frans J. Van Meirb, and André A. Weynsa
a Laboratory of Veterinary Anatomy and Embryology, Faculty of Medicine, University of Antwerp, Antwerp, Belgium
b Laboratory of Cell Biology and Histology, Faculty of Medicine, University of Antwerp, Antwerp, Belgium

Correspondence to: Christa J. Van Ginneken, Lab. of Veterinary Anatomy and Embryology, Slachthuislaan 68, 2060 Antwerp, Belgium..

Methods that visualize subsets as well as the entire enteric neuron population are not readily available or have proved to be unreliable. Therefore, we attempted to combine NADPH-d histochemistry, AChE histochemistry, and CGRP immunohistochemistry, techniques that mark subsets of enteric neurons, with a technique that appeared to visualize the entire enteric neuron population, the cuprolinic blue staining method. To guarantee representative staining results, the individual staining methods were modified by using microwaves. In addition, this preserved the characteristics of each of the individual techniques. The distribution of NADPH-d, AChE, and CGRP corresponded well 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 visualize the total population as well as subsets of enteric neurons. (J Histochem Cytochem 47:13–21, 1999)

Key Words: enteric nervous system, cuprolinic blue, NADPH-d, acetylcholinesterase, CGRP, microwave, double labeling


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