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Flow Cytometry and Mass Cytometry for Measuring the Immune Cell Infiltrate in Atherosclerotic Arteries

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Atherosclerosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2419))

Abstract

Atherosclerosis is characterized by the abundant infiltration of immune cells starting at early stages and progressing to late stages of the disease. The study and characterization of immune cells infiltrating and residing in the aorta has being tackled by several methodologies such as flow cytometry and mass cytometry (CyTOF). Flow cytometry has been primarily used to address the aortic leukocyte composition; however, only a limited number of markers can be analyzed simultaneously. CyTOF started to overcome these limitations by employing rare element-tagged antibodies and combines mass spectrometry with the ease and precision of flow cytometry. CyTOF currently allows for the simultaneous measurement of more than 40 cellular parameters at single-cell resolution.

In this chapter, we describe the methodology used to isolate single immune cells from mouse aortas, followed by protocols for flow cytometry and CyTOF for aortic immune cell characterization.

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Acknowledgments

This work was supported by grants to M.O. from the American Heart Association (AHA18POST34060251), and from The Conrad Prebys Foundation Award. M.A.M acknowledges support from the American Cancer Society Postdoctoral Fellowship (PF-20-132-01-LIB) and the National Institute of Health (2T32AR064194). C. C. H. Acknowledges support from NIH grants U01 CA224766, R01 CA202987, R01 HL134236, and P01 HL136275. K. L. was supported by grants NIH HL 115232, 145241, and HL088093.

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Author notes

  1. Marco Orecchioni and Melissa A. Meyer contributed equally to this work.

Authors and Affiliations

  1. La Jolla Institute for Immunology, La Jolla, CA, USA

    Marco Orecchioni, Melissa A. Meyer, Catherine C. Hedrick & Klaus Ley

  2. Department of Bioengineering, University of California San Diego, La Jolla, CA, USA

    Klaus Ley

Authors
  1. Marco Orecchioni
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  2. Melissa A. Meyer
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  3. Catherine C. Hedrick
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  4. Klaus Ley
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Corresponding authors

Correspondence to Catherine C. Hedrick or Klaus Ley .

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Editors and Affiliations

  1. School of Biosciences, Cardiff University, Cardiff, UK

    Dipak Ramji

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Orecchioni, M., Meyer, M.A., Hedrick, C.C., Ley, K. (2022). Flow Cytometry and Mass Cytometry for Measuring the Immune Cell Infiltrate in Atherosclerotic Arteries. In: Ramji, D. (eds) Atherosclerosis. Methods in Molecular Biology, vol 2419. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1924-7_47

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  • DOI: https://doi.org/10.1007/978-1-0716-1924-7_47

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1923-0

  • Online ISBN: 978-1-0716-1924-7

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