Redefining Manual White Blood Cell Analysis: A Rapid and Simplified 3x3 Grid Method Using Leishman Stained Peripheral Smears- A Pilot Study

Background: The conventional Battlemant method used for differential leucocyte count evaluation is time consuming and labour intensive. Lack of uniform distribution of leukocytes across the central and peripheral areas of the smear prompted us to explore a grid-based method to identify the best sites for differential leucocyte count evaluation. Aim: To evaluate white blood cells distribution in central and peripheral regions of blood smears and to assess the feasibility of a standardized 3 x 3 field-based method. Methods: 1x1 mm grids were marked at three sites each on peripheral and central region of Leishman stained blood smear (total 6). Three groups were classified across the grids. Group 1: 3 grids at periphery; Group 2: 3 grids at centre; Group 3: two central and one peripheral. Around 100 white blood cells were morphologically categorized in each group. Results and Statistics: Group 2 yielded the highest mean total leucocyte count (157.50 +- 65.95), followed by the Group 3: 147.80 +- 65.83. Basophils were frequent in the group 1(0.73%) than in the group 2 (0.45%) or group 3 (0.51%). Monocytes showed a group 1 preponderance versus the group 2 (2.49% vs. 1.58-1.96%). The mean distorted cells were 4.40 +- 3.24 in group 1; 3.17 +- 2.61 in group 2 and 4.17 +- 3.14 in group 3. Conclusion: Maximum white blood cell count was in group 2 (centre). Occurrence of basophils at group 1 (periphery) was statistically significant (p=0.023) while distribution of other white blood cells was not significant. This validates group 3 (combination) grids to be reliable for manual white blood cell analysis.