VALIDATION OF AN AUTOMATED METHOD FOR LEUKOCYTE COUNTING IN HEMATOPOIETIC PROGENITOR CELL PRODUCTS USING THE ABX MICROS 60 ANALYZER
- 1Grupo Pulsa São Paulo, São Paulo, SP, Brazil
Abstract
Introduction: Leukocyte counting in hematopoietic progenitor cell (HPC) products is a critical component of quality control in cellular therapy and hematopoietic stem cell transplantation. Accurate cell quantification is essential to ensure product safety, process standardization, and compliance with quality requirements. Traditionally, leukocyte counts are determined manually using a Neubauer chamber, a method that is labor-intensive, time-consuming, and subject to interobserver variability. Furthermore, the reliability of the analysis depends heavily on operator expertise. Laboratory automation has emerged as a promising alternative, providing greater analytical standardization, shorter turnaround times, and improved reproducibility. Therefore, this study aimed to validate an automated leukocyte counting method for HPC products using the ABX Micros 60 analyzer and compare its performance with the conventional manual approach. Methods: Thirty HPC samples derived from peripheral blood (PB-HPC) and bone marrow (BM-HPC) were included in this study. Samples were obtained either by apheresis or iliac crest aspiration performed in the operating room. Manual leukocyte counts were performed in duplicate using a Neubauer chamber according to the standardized procedures established in our cell-processing laboratory. Simultaneously, the same samples were analyzed in duplicate using the ABX Micros 60 analyzer following a 1:10 dilution in Minidil reagent. Mean values, standard deviations, and coefficients of variation were calculated for all samples. The predefined acceptance criteria were as follows: (1) a variation of ≤ 10% between duplicate automated measurements in at least 90% of samples and (2) a variation of ≤ 20% between manual and automated measurements in at least 90% of samples. Results: Duplicate analyses performed using the automated analyzer demonstrated complete agreement among all evaluated samples, with 100% of measurements showing a variation of ≤ 10%, thereby confirming the excellent reproducibility of the method. Comparison between manual and automated methodologies showed that 28 of the 30 samples (93.3%) exhibited a variation of ≤ 20%, fulfilling the predefined validation criteria. Only two samples (6.7%) presented variations exceeding the established threshold. Overall, the results demonstrated the analytical stability and satisfactory performance of the automated method for leukocyte quantification in diluted HPC samples obtained from both peripheral blood and bone marrow sources. Additionally, the automated approach significantly improved processing efficiency and standardization compared with manual counting procedures. Conclusion: The automated leukocyte counting method using the ABX Micros 60 analyzer demonstrated adequate performance for the evaluation of hematopoietic progenitor cell products, showing high levels of precision, stability, reproducibility, and operational efficiency. The methodology successfully met all predefined validation criteria and proved to be a reliable alternative to conventional manual counting, with the potential to optimize laboratory workflows and strengthen quality assurance processes in cellular therapy laboratories.
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BibTeX
@article{calheiros2026validation,
title = {{VALIDATION OF AN AUTOMATED METHOD FOR LEUKOCYTE COUNTING IN HEMATOPOIETIC PROGENITOR CELL PRODUCTS USING THE ABX MICROS 60 ANALYZER}},
author = {Waldely Veneziani CALHEIROS and Miria Lemos DA SILVA and Nayanna Faria DE CARVALHO and Erica Mioto Braz MERCHAN and Mirianceli Coelho MENDONçA and Cesar De Almeida NETO},
journal = {Hematology, Transfusion and Cell Therapy},
year = {2026},
month = oct,
doi = {10.1016/j.htct.2026.106922},
url = {https://doi.org/10.1016/j.htct.2026.106922},
note = {CC BY},
}
RIS
TY - JOUR
TI - VALIDATION OF AN AUTOMATED METHOD FOR LEUKOCYTE COUNTING IN HEMATOPOIETIC PROGENITOR CELL PRODUCTS USING THE ABX MICROS 60 ANALYZER
AU - Waldely Veneziani CALHEIROS
AU - Miria Lemos DA SILVA
AU - Nayanna Faria DE CARVALHO
AU - Erica Mioto Braz MERCHAN
AU - Mirianceli Coelho MENDONçA
AU - Cesar De Almeida NETO
PY - 2026
DA - 2026/10/23
JO - Hematology, Transfusion and Cell Therapy
DO - 10.1016/j.htct.2026.106922
UR - https://doi.org/10.1016/j.htct.2026.106922
AB - Introduction: Leukocyte counting in hematopoietic progenitor cell (HPC) products is a critical component of quality control in cellular therapy and hematopoietic stem cell transplantation. Accurate cell quantification is essential to ensure product safety, process standardization, and compliance with quality requirements. Traditionally, leukocyte counts are determined manually using a Neubauer chamber, a method that is labor-intensive, time-consuming, and subject to interobserver variability. Furthermore, the reliability of the analysis depends heavily on operator expertise. Laboratory automation has emerged as a promising alternative, providing greater analytical standardization, shorter turnaround times, and improved reproducibility. Therefore, this study aimed to validate an automated leukocyte counting method for HPC products using the ABX Micros 60 analyzer and compare its performance with the conventional manual approach. Methods: Thirty HPC samples derived from peripheral blood (PB-HPC) and bone marrow (BM-HPC) were included in this study. Samples were obtained either by apheresis or iliac crest aspiration performed in the operating room. Manual leukocyte counts were performed in duplicate using a Neubauer chamber according to the standardized procedures established in our cell-processing laboratory. Simultaneously, the same samples were analyzed in duplicate using the ABX Micros 60 analyzer following a 1:10 dilution in Minidil reagent. Mean values, standard deviations, and coefficients of variation were calculated for all samples. The predefined acceptance criteria were as follows: (1) a variation of ≤ 10% between duplicate automated measurements in at least 90% of samples and (2) a variation of ≤ 20% between manual and automated measurements in at least 90% of samples. Results: Duplicate analyses performed using the automated analyzer demonstrated complete agreement among all evaluated samples, with 100% of measurements showing a variation of ≤ 10%, thereby confirming the excellent reproducibility of the method. Comparison between manual and automated methodologies showed that 28 of the 30 samples (93.3%) exhibited a variation of ≤ 20%, fulfilling the predefined validation criteria. Only two samples (6.7%) presented variations exceeding the established threshold. Overall, the results demonstrated the analytical stability and satisfactory performance of the automated method for leukocyte quantification in diluted HPC samples obtained from both peripheral blood and bone marrow sources. Additionally, the automated approach significantly improved processing efficiency and standardization compared with manual counting procedures. Conclusion: The automated leukocyte counting method using the ABX Micros 60 analyzer demonstrated adequate performance for the evaluation of hematopoietic progenitor cell products, showing high levels of precision, stability, reproducibility, and operational efficiency. The methodology successfully met all predefined validation criteria and proved to be a reliable alternative to conventional manual counting, with the potential to optimize laboratory workflows and strengthen quality assurance processes in cellular therapy laboratories.
SN - 2531-1379
SN - 2531-1387
ER -