Immunophenotyping of Posttraumatic Neutrophils on a Routine Haematology Analyser

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.

Neutrophil CD64 expression as a longitudinal biomarker for severe disease and acute infection in critically ill patients

“…The hematology analyzer uses spectrophotometry, electrical impedance, laser light scattering [multi‐angle polarized scatter separation (MAPPS)], and 3 color fluorescent technologies to classify blood cells . Moreover, the analyzer is equipped with an integrated fluorescence (488 blue diode) laser and three fluorescent detectors, which allows using as a flow cytometer in a routine laboratory setting . The results are reported as a PMN CD64 index.…”

A single preoperative blood test predicts postoperative sepsis and pneumonia after coronary bypass or open aneurysm surgery

“…In the University Medical Center Utrecht (UMCU), all results are stored in the Utrecht Patient Oriented Database (UPOD). All haematology measurements used in this study were measured with the CELL‐DYN Sapphire (Abbott Diagnostics, Santa‐Clara, CA, USA), which is an automated multi‐parameter haematology analyzer for clinical purposes . The CELL‐DYN Sapphire uses laser light scattering, spectrophotometry and electrical impedance to describe distinct morphological traits (cell size, shape and granularity) of red blood cells, platelets and white blood cells.…”

“…All haematology measurements used in this study were measured with the CELL-DYN Sapphire (Abbott Diagnostics, Santa-Clara, CA, USA), which is an automated multi-parameter haematology analyzer for clinical purposes. 13,14 The CELL-DYN Sapphire uses laser light scattering, spectrophotometry and electrical impedance to describe distinct morphological traits (cell size, shape and granularity) of red blood cells, platelets and white blood cells. An example of an overlay of two erythrocyte volume distribution curves is presented in Figure 1; both patients demonstrated comparable MCV but different RDW.…”

Activation State of the Circulating Neutrophil in Isolated Chest Trauma Patients: Characterization of Surface Receptor Expression

“…Trauma is a major cause of morbidity and mortality which are mainly due to a dysfunctional in the immune system [1,2]. The acute respiratory distress syndrome (ARDS) is a common complication after severe injury, the incidence of ARDS is found to be low in patients with isolated chest injury [3] and with increased severity of pulmonary contusion the incidence increase [4][5][6].…”