ClearLLab 10C reagents panel can be applied to analyze paucicellular samples by flow cytometry

Area of Science:

• Hematology
• Immunophenotyping
• Clinical Diagnostics

Background:

• The FDA-approved ClearLLab 10C Reagents Panel simplifies leukemia and lymphoma diagnosis via flow cytometry.
• A limitation is its requirement for 3 × 10⁶ cells/mL, which is unachievable for paucicellular samples.

Purpose of the Study:

• To evaluate the reliability of the ClearLLab 10C 10-color panel for analyzing specimens with low cell concentrations.
• To determine if the panel can be effectively used with cell concentrations significantly lower than the FDA-approved standard.

Main Methods:

• Serial dilutions of 16 samples (5 normal, 11 abnormal) were prepared, ranging from 3.0 × 10⁶ to 0.0469 × 10⁶ cells/mL.
• Samples were stained using the B-cell and T-cell panels of the ClearLLab 10C system.
• Mean fluorescence intensity (MFI) was measured, and deviations from the standard concentration were calculated and analyzed using Bland-Altman, correlation, and regression methods.

Main Results:

• Antigen expression patterns remained consistent across all tested cell concentrations.
• The mean deviation of MFI from the standard concentration never exceeded 10% for any antibody.
• Bland-Altman analysis showed similarity between standard and diluted cell suspension results, with high positive correlation.
• A 100% concordance in antigen expression patterns was observed between the FDA-approved concentration and the lowest cell density tested.

Conclusions:

• The ClearLLab 10C panel reliably determines marker expression for leukemias and lymphomas in paucicellular samples.
• The panel is effective with cell concentrations as low as 0.0469 × 10⁶ cells/mL (64-fold lower than FDA-approved).
• This finding significantly broadens the clinical applicability of the ClearLLab 10C platform for flow cytometry diagnostics.