Optimized workflow for human PBMC multiomic immunosurveillance studies

Palak C Genge1, Charles R Roll1, Alexander T Heubeck1

  • 1Allen Institute for Immunology, 615 Westlake Avenue N, Seattle, WA 98109, USA.

STAR Protocols
|November 22, 2021
PubMed

Insights

This study presents a robust pipeline for deep immune profiling of human peripheral blood mononuclear cells. The protocol minimizes batch effects, ensuring reproducible data for both flow cytometry and single-cell RNA-seq analyses.

Area of Science:

  • Immunology
  • Molecular Biology
  • Genomics

Background:

  • Deep immune profiling is crucial for understanding human health and disease.
  • Interpreting complex immunological data necessitates consistent laboratory processing to minimize batch-to-batch variation.

Purpose of the Study:

  • To detail a robust pipeline for the comprehensive profiling of human peripheral blood mononuclear cells.
  • To reduce batch effects and enhance data reproducibility in high-dimensional flow cytometry and single-cell RNA-seq.

Main Methods:

  • Development of a standardized laboratory pipeline for processing human peripheral blood mononuclear cells.
  • Application of high-dimensional flow cytometry for immune cell surface marker analysis.
  • Implementation of single-cell RNA sequencing for transcriptomic profiling.

Main Results:

  • The developed protocols significantly reduce batch effects in immunological data.
  • The pipeline generates reproducible data, enhancing the reliability of experimental findings.
  • Increased throughput was achieved, allowing for more extensive immune profiling studies.

Conclusions:

  • This robust pipeline provides a standardized approach for deep immune profiling.
  • The methods ensure data consistency and reproducibility, crucial for biological interpretation.
  • The protocol is essential for advancing the understanding of the human immune system in various conditions.

Related Concept Videos