Optimum Volume of Water to Be Added to the Leishman Stain While Preparing a Blood Smear
Background and Aim: During blood smear preparation, Leishman stain is first added to the slide for fixation. Then, aqueous fluid (e.g., distilled water [DW] and buffered water) is added and mixed with the stain. Different books suggest different amount of aqueous fluid to add with the stain. With this background, the aim of this study was to compare the quality of staining of blood smears stained by Leishman stain with different amount of DW. Methods: A total of 4 glass slides, with blood smear were stained by Leishman stain with different amount of DW – half the amount, equal the amount, 1½ times the amount, and double the amount of stain. After washing and drying, these blood films were seen under microscope, and the quality of staining was rated by 10 observers on 10‑point scale. Scores for smears were compared by one‑way ANOVA in Microsoft Excel 2010. Results: The mean score for blood film stained with half DW was 7.5 ± 0.71, double DW was 7.9 ± 0.74, 1½ times DW was 7.7 ± 0.67, and double DW was 7.6 ± 0.52. There was no difference in mean score (P = 0.58) among the films when tested by ANOVA. Conclusion: There was no difference in quality of staining among smears stained by Leishman stain with different amount of DW. Hence, any volume of DW ranging from half the amount to double the amount of stain may be suggested for staining. However, half the amount may be avoided due to chances of stain precipitation.