Optimized workflow for human PBMC multiomic immunosurveillance studies
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Updated: Oct 12, 2025

Optimized Workflow for Iterative Bleaching Extends Multiplexity Imaging of Highly Autofluorescent Clinical Samples
Published on: July 11, 2025
Palak C Genge1, Charles R Roll1, Alexander T Heubeck1
1Allen Institute for Immunology, 615 Westlake Avenue N, Seattle, WA 98109, USA.
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).

