Reflection contrast microscopy within chrome-alum haematoxylin stained thick tissue-sections.

This paper introduces two innovations in reflection contrast microscopy (RCM): (1) an extended application for qualitative light microscopic investigations; and (2) a novel method for quantification in cytochemistry. (1) We found out that RCM cannot only be used for surface characterizations and in thin sections but also within thick tissue-sections. The use of the RCM technique is demonstrated on slides of the supraoptic nucleus (SON) of the rat stained with chrome-alum haematoxylin: Among all the stained structures only neurosecretory granules are found to cause reflections. The visualization of the neurosecretion and its distribution is more distinct and of sharper contrast than in bright field microscopy. (2) The improved differentiation allows the quantification of neurosecretion in tissue-sections by combining RCM with grey-scale image analysis.