Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Flow cytometry (FC) is a critical tool in immunology for analyzing immune cells.
• Inconsistent application of FC techniques can lead to unreliable immunological data.
• A need exists for standardized guidelines to ensure accuracy and reproducibility in FC experiments.

Purpose of the Study:

• To establish consensus guidelines for the effective use of flow cytometry in immunology.
• To provide practical advice and theoretical background for immunologists utilizing FC.
• To minimize common errors and enhance the quality of immunological data derived from FC.

Main Methods:

• Consensus-based development involving experts in immunology and flow cytometry.
• Compilation of theoretical principles and practical aspects of FC techniques.
• Inclusion of detailed immune cell phenotyping for murine and human samples.
• Description of the latest FC applications and data analysis strategies.

Main Results:

• Comprehensive guidelines covering major immune cell types and their phenotypes.
• Detailed tables of murine and human immune cell phenotypes.
• Explanation of current FC techniques, applications, and data analysis.
• Inclusion of expert tips, tricks, and common pitfalls to avoid.

Conclusions:

• These guidelines serve as an essential resource for immunologists using flow cytometry.
• Adherence to these guidelines will improve the reliability and reproducibility of immunological research.
• The document empowers researchers to avoid common errors and optimize FC data generation and analysis.