Original article / research
A Comparative Study between Conventional Leishman Stain and a Modified Blood Stain for the Evaluation of Haematologic Elements
Dr. Ashida M Krishnan,
Associate Professor (non cadre), Department of Pathology, Government Medical College, Thiruvananthapuram-695011, Kerala, India.
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Introduction: Leishman is the most common Romanowsky stain globally used for studying peripheral blood morphology. Leishman stain is a compound neutral dye using combination of either eosin Y or eosin B with partially oxidised methylene blue. Inconsistent staining reactions occurring with these dyes are attributed to the difficulty in controlling the methylene blue oxidation process.
Aim: To compare the staining qualities of Leishman with a modified blood stain called Villanueva stain which has a combination of non-oxidised methylene blue, azure II and Eosin.
Materials and Methods: Present cross-sectional study was carried out in Central Haematology laboratory of a tertiary health care centre in Southern India, from August 2017 to January 2018. Blood Sample was collected in anticoagulated vials. Two thin wedge smears were prepared from each sample and one was stained with Villanueva and other with Leishman stain. The staining characteristics were assessed and scored in terms of 5 parameters- nuclear chromatin, eosinophil granules, neutrophil granules, platelets, Red Blood Cell (RBC) staining and finally all scores were summed as poor/satisfactory/good/excellent by an experienced pathologist. The results were entered in Microsoft excel sheet and were analysed by Statistical Package for the Social Sciences (SPSS) software version 16.0.
Results: The chromatin staining was superior in Leishman staining (Measure agreement kappa = 0.028, p=0.631, McNemar test p=0.001). For platelet staining and RBC the statistical agreement between both tests were moderate and for neutrophils and eosinophil granules agreement was poor between two staining methods.
Conclusion: The modified stain can be easily prepared from chemicals cheaply available. Though modified stain gave inferior results with conventional Leishman stain, present study was helpful to know if peripheral smear can be interpreted with this modified stain in case of Leishman stain shortage. Alternative staining method deserves importance in this era of global lock down where manufacture and transportation of chemicals are adversely affected worldwide.
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