Functional analysis of human circulating immune cells based on high-dimensional mass cytometry

Xiuxing Liu1, Jianjie Lv1, Huishi Wang1

  • 1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, China.

STAR Protocols
|April 18, 2022
PubMed

Insights

This study introduces a new mass cytometry workflow to reduce batch effects in clinical immune system analysis. The validated method enhances data reliability for complex blood samples.

Area of Science:

  • Immunology
  • Biotechnology
  • Clinical Diagnostics

Background:

  • Mass cytometry offers high-resolution, high-dimensional analysis of complex biological systems like the human immune system.
  • Clinical applications of mass cytometry are hindered by batch effects arising from long-term sample collection, potentially masking true biological signals.
  • Standardized and validated workflows are crucial for reliable clinical implementation of advanced analytical techniques.

Purpose of the Study:

  • To present a validated and streamlined mass cytometry workflow optimized for clinical settings.
  • To address and mitigate the issue of batch effects in the analysis of clinical immune samples.
  • To enhance the reliability and applicability of mass cytometry in clinical research and diagnostics.

Main Methods:

  • Development of a mass cytometry workflow incorporating fixed staining protocols suitable for clinical use.
  • Implementation of optimized cellular barcode staining patterns to improve sample multiplexing and data quality.
  • Validation of the workflow's efficacy in reducing batch effects and ensuring data consistency.

Main Results:

  • The proposed workflow effectively reduces batch effects commonly encountered in long-term clinical sample collection.
  • Fixed staining and optimized barcode patterns contribute to more robust and reproducible mass cytometry data.
  • The workflow demonstrates improved reliability for analyzing complex immune system profiles in clinical settings.

Conclusions:

  • The presented mass cytometry workflow provides a validated solution for overcoming batch effect challenges in clinical sample analysis.
  • This streamlined approach extends the practical application range and leverages the full advantages of mass cytometry in clinical research.
  • The workflow enhances the potential for accurate immune system profiling and biomarker discovery in clinical settings.