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Provides a Computational Framework for the Nonspecialist to Profile High-dimensional Cytometry Data

Abstract

High-dimensional cytometry is an innovative tool for immune monitoring in health and disease, and it has provided novel insight into the underlying biology as well as biomarkers for a variety of diseases. However, the analysis of large multiparametric datasets usually requires specialist computational knowledge. Here, we describe (https://github.com/kordastilab/ImmunoCluster), an R package for immune profiling cellular heterogeneity in high-dimensional liquid and imaging mass cytometry, and flow cytometry data, designed to facilitate computational analysis by a nonspecialist. The analysis framework implemented within is readily scalable to millions of cells and provides a variety of visualization and analytical approaches, as well as a rich array of plotting tools that can be tailored to users' needs. The protocol consists of three core computational stages: (1) data import and quality control; (2) dimensionality reduction and unsupervised clustering; and (3) annotation and differential testing, all contained within an R-based open-source framework.

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References
1.
Angerer P, Haghverdi L, Buttner M, Theis F, Marr C, Buettner F . destiny: diffusion maps for large-scale single-cell data in R. Bioinformatics. 2015; 32(8):1241-3. DOI: 10.1093/bioinformatics/btv715. View

2.
Ashhurst T, Marsh-Wakefield F, Putri G, Spiteri A, Shinko D, Read M . Integration, exploration, and analysis of high-dimensional single-cell cytometry data using Spectre. Cytometry A. 2021; 101(3):237-253. DOI: 10.1002/cyto.a.24350. View

3.
Ashhurst T, Cox D, Smith A, King N . Analysis of the Murine Bone Marrow Hematopoietic System Using Mass and Flow Cytometry. Methods Mol Biol. 2019; 1989:159-192. DOI: 10.1007/978-1-4939-9454-0_12. View

4.
DiGiuseppe J, Cardinali J, Rezuke W, Peer D . PhenoGraph and viSNE facilitate the identification of abnormal T-cell populations in routine clinical flow cytometric data. Cytometry B Clin Cytom. 2017; 94(5):588-601. PMC: 5834343. DOI: 10.1002/cyto.b.21588. View

5.
Hartmann F, Babdor J, Gherardini P, Amir E, Jones K, Sahaf B . Comprehensive Immune Monitoring of Clinical Trials to Advance Human Immunotherapy. Cell Rep. 2019; 28(3):819-831.e4. PMC: 6656694. DOI: 10.1016/j.celrep.2019.06.049. View