Quantitative Flow Cytometry: Concerns and Recommendations in Clinic and Research

Area of Science:

• Clinical diagnostics
• Cellular analysis
• Biotechnology

Background:

• Quantitative flow cytometry (QFCM) is evolving as a crucial tool for clinical diagnostics.
• Advancements in FC technology enable antigen quantification via median fluorescence intensity (MFI), making it clinically relevant.

Purpose of the Study:

• To highlight key concerns in quantitative FC methodology.
• To propose solutions for achieving uniform and standardized QFCM tests based on MFI.

Main Methods:

• Standardization relies on sample/reagent handling, instrument maintenance, and data analysis.
• Specialized beads are used for routine calibration to reduce inter-test variability.
• Agreed biological controls further minimize test variability.

Main Results:

• Inter- and intra-laboratory variations are influenced by several parameters.
• Routine calibration with specialized beads reduces variability between operators and instruments.
• Standardized biological controls significantly lower test variability.

Conclusions:

• QFCM plays a vital role in the clinical diagnostic toolbox.
• Recommendations can enhance the stability and accuracy of QFCM.
• Improved standardization ensures more reliable cell analyses.