Reporting an Experience: Improving the Feulgen Staining Technique for Better Visualizing of Nucleus

Noushin Jalayer Naderi

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URL :   http://research.shahed.ac.ir/WSR/WebPages/Report/PaperView.aspx?PaperID=84815
Date :  2018/02/07
Publish in :    Iranian Journal of Pathology
DOI :  https://doi.org/10.30699/ijp.13.1.106

Keywords : Feulgen S taining T echnique

Abstract :

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M icronuclei (MN) are fragments of chromosomes that fail to encompass into the nucleus and remains in the cytoplasm in cell division course (1). Monitoring MN in oral exfoliated epithelial cells for detecting exposed individuals to genotoxic agents was proposed by Stich et al. in 1982 for the first time (2). The genotoxic agents of tobacco, waterpipe and even environmental contaminants are recognized as initiator factors in producing the aberrations that results in MN production (3 - 5). Assessing the frequency of MN in oral exfoliated cells is a non - invasive reliable indicator of genotoxic da mage in human (6). Among different staining methods to demonstrate the nuclear abnormalities, Feulgen is one of the most reliable method. The Feulgen staining is a specific and sensitive method for evaluating DNA damages . It has been shown that using non - DNA specific stains to monitor nuclear anomalies lead to false - positive or false - negative results (7). In a routine Feulgen staining technique, slides are immersed in 5 mol/L HCl for 15 minutes, rinsed with distilled water for 3 minutes, stained with S chiffs reagent for 90 minutes, washed for 10 minutes and finally stained with 1 light green for 15 minutes (7 - 8). Using the Feulgen staining, nucleus and micronuclei appear magenta and cytoplasm seems green. From self - experience, i mmersing the stained slides in hematoxylin for 3 - 5 minutes and eosin for few seconds in the final step after staining the slides with light green, provides a greater ground contrast and very better visualizing of nucleus (Fig1).