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Mediators of Inflammation
Volume 2012 (2012), Article ID 509513, 6 pages
http://dx.doi.org/10.1155/2012/509513
Research Article

Immunophenotyping of Posttraumatic Neutrophils on a Routine Haematology Analyser

1Department of Trauma Surgery, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands
2Department of Respiratory Medicine, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands
3Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands

Received 1 October 2011; Accepted 6 December 2011

Academic Editor: Sascha Flohe

Copyright © 2012 Kathelijne Maaike Groeneveld et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Introduction. Flow cytometry markers have been proposed as useful predictors for the occurrence of posttraumatic inflammatory complications. However, currently the need for a dedicated laboratory and the labour-intensive analytical procedures make these markers less suitable for clinical practice. We tested an approach to overcome these limitations. Material and Methods. Neutrophils of healthy donors were incubated with antibodies commonly used in trauma research: CD11b (MAC-1), L-selectin (CD62L), FcγRIII (CD16), and FcγRII (CD32) in active form (MoPhab A27). Flow cytometric analysis was performed both on a FACSCalibur, a standard flow cytometer, and on a Cell-Dyn Sapphire, a routine haematology analyser. Results. There was a high level of agreement between the two types of analysers, with 41% for FcγRIII, 80% for L-selectin, 98% for CD11b, and even a 100% agreement for active FcγRII. Moreover, analysis on the routine haematology analyser was possible in less than a quarter of the time in comparison to the flow cytometer. Conclusion. Analysis of neutrophil phenotype on the Cell-Dyn Sapphire leads to the same conclusion compared to a standard flow cytometer. The markedly reduced time necessary for analysis and reduced labour intensity constitutes a step forward in implementation of this type of analysis in clinical diagnostics in trauma research.