Performance of spectral flow cytometry and mass cytometry for the study of innate myeloid cell populations

Frontiers in Immunology

Immunology

Sara Kassem¹  Kyra van der Pan¹  Brigitta A.E. Naber¹  Indu Khatri¹  Suzanne E.T. Comans¹  Alesha Louis¹  Anniek L. de Jager¹  Inge F. de Laat¹  Paula Díez²  Jacques J.M. van Dongen³  Cristina Teodosio³  Marjolijn Hameetman⁴  Alberto Orfao⁵ 

[1] Department of Immunology, Leiden University Medical Center (LUMC), Leiden, Netherlands;Department of Immunology, Leiden University Medical Center (LUMC), Leiden, Netherlands;Sarcomas and Experimental Therapeutics Laboratory, Health Research Institute of Asturias (ISPA) and Asturias Central University Hospital (HUCA), Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Oviedo, Asturias, Spain;Department of Immunology, Leiden University Medical Center (LUMC), Leiden, Netherlands;Translational and Clinical Research Program, Cancer Research Center (IBMCC; University of Salamanca - CSIC), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca and Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain;Flow Cytometry Core Facility, Leiden University Medical Center (LUMC), Leiden, Netherlands;Translational and Clinical Research Program, Cancer Research Center (IBMCC; University of Salamanca - CSIC), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca and Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain;

关键词: spectral flow cytometry;    myeloid cells;    immunophenotyping;    cyTOF;    mass cytometry;   

DOI  :  10.3389/fimmu.2023.1191992

 received in 2023-03-22, accepted in 2023-05-05,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionMonitoring of innate myeloid cells (IMC) is broadly applied in basic and translational research, as well as in diagnostic patient care. Due to their immunophenotypic heterogeneity and biological plasticity, analysis of IMC populations typically requires large panels of markers. Currently, two cytometry-based techniques allow for the simultaneous detection of ≥40 markers: spectral flow cytometry (SFC) and mass cytometry (MC). However, little is known about the comparability of SFC and MC in studying IMC populations.MethodsWe evaluated the performance of two SFC and MC panels, which contained 21 common markers, for the identification and subsetting of blood IMC populations. Based on unsupervised clustering analysis, we systematically identified 24 leukocyte populations, including 21 IMC subsets, regardless of the cytometry technique.ResultsOverall, comparable results were observed between the two technologies regarding the relative distribution of these cell populations and the staining resolution of individual markers (Pearson’s ρ=0.99 and 0.55, respectively). However, minor differences were observed between the two techniques regarding intra-measurement variability (median coefficient of variation of 42.5% vs. 68.0% in SFC and MC, respectively; p<0.0001) and reproducibility, which were most likely due to the significantly longer acquisition times (median 16 min vs. 159 min) and lower recovery rates (median 53.1% vs. 26.8%) associated with SFC vs. MC.DiscussionAltogether, our results show a good correlation between SFC and MC for the identification, enumeration and characterization of IMC in blood, based on large panels (>20) of antibody reagents.

【 授权许可】

Unknown   
Copyright © 2023 van der Pan, Khatri, de Jager, Louis, Kassem, Naber, de Laat, Hameetman, Comans, Orfao, van Dongen, Díez and Teodosio