Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood...

Area of Science:

• Immunology
• Biotechnology
• Clinical Research

Background:

• Multi-parametric flow cytometry is crucial for immune cell analysis in health and disease.
• Standardized procedures are vital for reliable comparisons in population-based and clinical studies.

Purpose of the Study:

• To develop and highlight standardized, semi-automated procedures for human whole blood immunophenotyping.
• To establish robust protocols for eight-color flow cytometry applicable to large-scale studies.

Main Methods:

• Optimization of eight-color antibody panels and staining/lysis procedures.
• Implementation of pre-analytic steps using a semi-automated robotic workflow.
• Development of four distinct panels for major immune cell populations (PMN, T, B, NK, monocytes, DC).

Main Results:

• Successful optimization of antibody panels and staining procedures.
• Implementation of a semi-automated workflow for pre-analytic sample handling.
• Development of four comprehensive panels for enumerating and phenotyping key immune cell subsets.

Conclusions:

• The developed protocols provide a foundation for defining reference values in healthy populations.
• The approach offers robust, affordable, semi-automated eight-color flow cytometry for population-based and clinical studies.
• Standardized immunophenotyping enhances the comparability and reliability of immune cell data.