Using Intravital Microscopy to Study the Role of MIF in Leukocyte Trafficking In Vivo

  • M. Ursula Norman
  • Michael J. HickeyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2080)


In vivo visualization of the microvasculature of the mouse cremaster muscle has been fruitful in the evaluation of the role of macrophage migration inhibitory factor in promotion of leukocyte trafficking. Here we explain how to undertake this preparation, including details on mouse anesthesia, securing intravenous access, and cremaster muscle exteriorization. We also provide information on the various microscopy modalities now available for imaging microvascular preparations of this nature.

Key words

Intravital microscopy Leukocyte interactions Rolling Adhesion Transmigration Cremaster muscle Postcapillary venule 



This work is supported by funding from the National Health and Medical Research Council of Australia (1042775, MJH) and the Monash University School of Clinical Sciences at Monash Health (MUN).


  1. 1.
    Calandra T, Roger T (2003) Macrophage migration inhibitory factor: a regulator of innate immunity. Nat Rev Immunol 3(10):791–800CrossRefGoogle Scholar
  2. 2.
    Ayoub S, Hickey MJ, Morand EF (2008) Mechanisms of disease: macrophage migration inhibitory factor in SLE, RA and atherosclerosis. Nat Clin Pract Rheumatol 4(2):98–105CrossRefGoogle Scholar
  3. 3.
    Bernhagen J et al (1996) An essential role for macrophage migration inhibitory factor in the tuberculin delayed-type hypersensitivity reaction. J Exp Med 183:277–282CrossRefGoogle Scholar
  4. 4.
    Lan HY et al (1997) The pathogenic role of macrophage migration inhibitory factor in immunologically induced kidney disease in the rat. J Exp Med 185(8):1455–1465CrossRefGoogle Scholar
  5. 5.
    Leech M et al (1998) Involvement of macrophage migration inhibitory factor in the evolution of rat adjuvant arthritis. Arthritis Rheum 41(5):910–917CrossRefGoogle Scholar
  6. 6.
    Hoi AY et al (2006) Macrophage migration inhibitory factor deficiency attenuates macrophage recruitment, glomerulonephritis, and lethality in MRL/lpr mice. J Immunol 177(8):5687–5696CrossRefGoogle Scholar
  7. 7.
    Gregory JL et al (2004) Reduced leukocyte-endothelial cell interactions in the inflamed microcirculation of macrophage migration inhibitory factor-deficient mice. Arthritis Rheum 50:3023–3034CrossRefGoogle Scholar
  8. 8.
    Gregory JL et al (2006) Macrophage migration inhibitory factor induces macrophage recruitment via CC chemokine ligand 2. J Immunol 177(11):8072–8079CrossRefGoogle Scholar
  9. 9.
    Bernhagen J et al (2007) MIF is a noncognate ligand of CXC chemokine receptors in inflammatory and atherogenic cell recruitment. Nat Med 13(5):587–596CrossRefGoogle Scholar
  10. 10.
    Stark K et al (2013) Capillary and arteriolar pericytes attract innate leukocytes exiting through venules and ‘instruct’ them with pattern-recognition and motility programs. Nat Immunol 14(1):41–51CrossRefGoogle Scholar
  11. 11.
    Ley K et al (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7(9):678–689CrossRefGoogle Scholar
  12. 12.
    Hickey MJ, Westhorpe CL (2013) Imaging inflammatory leukocyte recruitment in kidney, lung and liver – challenges to the multi-step paradigm. Immunol Cell Biol 91(4):281–289CrossRefGoogle Scholar
  13. 13.
    Cheng Q et al (2010) Macrophage migration inhibitory factor increases leukocyte-endothelial interactions in human endothelial cells via promotion of expression of adhesion molecules. J Immunol 185(2):1238–1247CrossRefGoogle Scholar
  14. 14.
    Santos LL et al (2011) Macrophage migration inhibitory factor regulates neutrophil chemotactic responses in inflammatory arthritis in mice. Arthritis Rheum 63(4):960–970CrossRefGoogle Scholar
  15. 15.
    Fan H et al (2011) Macrophage migration inhibitory factor and CD74 regulate macrophage chemotactic responses via MAPK and Rho GTPase. J Immunol 186(8):4915–4924CrossRefGoogle Scholar
  16. 16.
    Wee JL et al (2015) Tetraspanin CD37 regulates b2 integrin-mediated adhesion and migration in neutrophils. J Immunol 195(12):5770–5779CrossRefGoogle Scholar
  17. 17.
    Nicholls AJ et al (2018) Activation of the sympathetic nervous system modulates neutrophil function. J Leukoc Biol 103(2):295–309CrossRefGoogle Scholar
  18. 18.
    Woodfin A et al (2011) The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo. Nat Immunol 12(8):761–769CrossRefGoogle Scholar
  19. 19.
    Vanheule V et al (2017) CXCL9-derived peptides differentially inhibit neutrophil migration in vivo through interference with glycosaminoglycan interactions. Front Immunol 8:530CrossRefGoogle Scholar
  20. 20.
    Mempel TR, Henrickson SE, von Andrian UH (2004) T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Nature 427:154–159CrossRefGoogle Scholar
  21. 21.
    Devi S et al (2013) Multiphoton imaging reveals a novel leukocyte recruitment paradigm in the glomerulus. Nat Med 19:107–112CrossRefGoogle Scholar
  22. 22.
    Snelgrove SL et al (2017) Activated renal dendritic cells cross present intrarenal antigens after ischemia reperfusion injury. Transplantation 101:1013–1024CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Centre for Inflammatory Diseases, Department of Medicine, School of Clinical Sciences at Monash Health, Faculty of Medicine, Nursing and Health Sciences, Monash Medical CentreMonash UniversityClaytonAustralia
  2. 2.Centre for Inflammatory Diseases, School of Clinical Sciences at Monash Health, Faculty of Medicine, Nursing and Health Sciences, Monash Medical CentreMonash UniversityClaytonAustralia

Personalised recommendations