Discussion

Background
Drug discovery and clinical research applications often require precise, longitudinal, multi-parametric and simultaneous phenotyping and analysis of human leukocyte populations. Recent advances in optics and sensors have enhanced the potential of multi-colour flow cytometry to be employed as the predominant technology for such purposes. However, inherent variabilities brought about by the need for complex reagent preparation requiring the addition of multiple antibody markers and obscure instrument setup requiring repeated compensation adjustments owing to tandem dye conjugate lot changes over time have restricted widespread application of flow cytometry. Beckman Coulter’s DuraCloneTM technology endeavors to overcome the limitation by ensuring standardization in the entire process work flow with the introduction of multicolor reagents in a dry, unitized format that is stable at room temperature, coupled with easy, automated compensation setup process.

Study
Four different multicolor reagent panels were designed for phenotyping and analysis of leukocyte subpopulations (DuraClone Basic Phenotyping Panel), B lymphocyte subpopulations (DuraClone IM B cells panel), T lymphocyte subsets (DuraClone IM T cells panel) and Dendritic Cells (DuraClone IM Dendritic cells panel). Leukocyte subpopulations of multiple donors were analyzed using the above mentioned multicolor panels. Compensation kits comprising of generic single conjugates for non-tandem dyes and panel specific conjugates for tandem dyes were used for generating automated compensation matrices using the NaviosTM flow cytometer’s Auto-Setup scheduler.

Results
Compensation kit along with Auto-Setup scheduler resulted in the generation of accurate compensation matrices which required minimal adjustments post sample acquisition. The panels produced precise and reproducible results on recruitment of each subpopulation of interest.

Conclusions
This study demonstrates the potential of DuraClone multicolor reagents to simplify and standardize the instrument setup and reagent workflow in multicolor flow cytometry applications. This will enable coherent and consistent monitoring of various immune cell phenotypes for research and clinical applications.