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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.
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.
Abstract:
Deep immune profiling is essential for understanding the human immune system in health and disease. Successful biological interpretation of this data requires consistent laboratory processing with minimal batch-to-batch variation. Here, we detail a robust pipeline for the profiling of human peripheral blood mononuclear cells by both high-dimensional flow cytometry and single-cell RNA-seq. These protocols reduce batch effects, generate reproducible data, and increase throughput. For complete details on the use and execution of this protocol, please refer to Savage et al. (2021).

