Implementation of monoclonal antibody fluorescence on the Abbott CELL-DYN Sapphire haematology analyser: evaluation of lymphoid, myeloid and platelet markers.

Apart from qualitative flags, that are typically inefficient and uninformative, haematology instruments provide little meaningful information about lymphocyte populations or the lineage of atypical or immature elements, The CELL-DYN Sapphire haematology analyser uses integrated optical and fluorescence (488 nm) measurements, with FL1 (FITC) and FL2 (PE) detectors being configured for fluorescent analysis. As monoclonal antibodies (Mab) are widely used as cellular probes, and are likely to constitute the future basis for immunodifferentials, we explored the feasibility of implementing immunofluorescence on this routine haematology analyser. An extensive series of Mab (CD2, CD3, CD4, CD8, CD11b, CD13, CD14, CD16, CD19, CD22, CD33, CD34, CD41, CD42b, CD45, CD56, CD61, CD64, CD235a and HLA-DR) were tested singly or in FITC/PE combinations. Analyser processing and data acquisition was achieved using CD-Sapphire automated CD61 immunoplatelet or CD3/4/8 assay procedures and, apart from mixing EDTA-blood and antibody, no further sample manipulation was required. Downloaded raw files were processed with cytometry software, and all evaluated reagents showed population discrimination analogous to flow cytometry. Practical procedures were straightforward and required minimal operator training. Extended information that can be obtained from monoclonal antibodies with a routine haematology analyser has the potential to extend haematology laboratory practices and positively impact laboratory and clinical efficiency.