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Crossover study of three methods for low-level white blood cell counting on two types of flow cytometer

Crossover study of three methods for low-level white blood cell counting on two types of flow cytometer

Cytometry. Part B, Clinical Cytometry 54(1): 39-45

Background: Flow cytometric methods were previously shown to be preferable to microscopic and volumetric methods for counting residual white blood cells (WBCs). In this study, three flow cytometric, low-level WBC counting methods were cross compared using two flow cytometers. Methods: Double-filtered red cell and platelet concentrates were spiked with different amounts of WBC to obtain panels of unspiked and 0.3, 1.0, 3.3, and 10.0 WBC/mul. The methods of BD Biosciences (BDB), Beckman-Coulter (BC), and an in-house method were performed on flow cytometers from BDB and BC. Samples were measured in ninefold. We required that (a) r2 be at least 0.98 (linearity), (b) at least 80% of observations fell within 20% of expected values (accuracy), and (c) the coefficients of variation be at least 20% (precision) for samples containing at least 3.3 WBC/mul. Results: For the red cell panel, our requirements were met by the BDB method on both flow cytometers and by the BC and in-house methods on the BDB flow cytometer only. For the platelet panel, our requirements were met on all combinations of methods and flow cytometers, except for the in-house method on the BDB flow cytometer. Intra-assay variation was lowest for the BDB method, irrespective of the type of flow cytometer used. Conclusion: Based on accuracy and precision, the BDB method on the BDB flow cytometer produced the best results for counting low-level WBCs.

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Accession: 010398042

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PMID: 12827666

DOI: 10.1002/cyto.b.10015

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