Advertisement

Microfluidic Platform to Quantify Neutrophil Migratory Decision-Making

  • Brittany P. Boribong
  • Amina Rahimi
  • Caroline N. JonesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1960)

Abstract

Neutrophils are the most abundant leukocytes in blood, serving as the first line of host defense in tissue damage and infections. Upon activation by chemokines released from pathogens or injured tissues, neutrophils migrate through complex tissue microenvironments toward sites of infections along the chemokine gradients, in a process named chemotaxis. However, current methods for measuring neutrophil chemotaxis require large volumes of blood and are often bulk, endpoint measurements. To address the need for rapid and robust assays, we engineered a novel dual gradient microfluidic platform that precisely quantifies neutrophil migratory decision-making with high temporal resolution. Here, we present a protocol to measure neutrophil migratory phenotypes (velocity, directionality) with single-cell resolution.

Keywords

Microfluidics Chemotaxis Immune cell Neutrophil migration phenotype Migratory decision-making 

References

  1. 1.
    Wu J, Wu X, Lin F (2013) Recent developments in microfluidics-based chemotaxis studies. Lab Chip 13(13):2484–2499.  https://doi.org/10.1039/c3lc50415hCrossRefPubMedGoogle Scholar
  2. 2.
    Irimia D, Ellett F (2016) Big insights from small volumes: deciphering complex leukocyte behaviors using microfluidics. J Leukoc Biol 100(2):291–304.  https://doi.org/10.1189/jlb.5RU0216-056RCrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Jones CN, Hoang AN, Martel JM et al (2016) Microfluidic assay for precise measurements of mouse, rat, and human neutrophil chemotaxis in whole-blood droplets. J Leukoc Biol 100(1):241–247.  https://doi.org/10.1189/jlb.5TA0715-310RRCrossRefPubMedGoogle Scholar
  4. 4.
    Hamza B, Irimia D (2015) Whole blood human neutrophil trafficking in a microfluidic model of infection and inflammation. Lab Chip 15(12):2625–2633.  https://doi.org/10.1039/c5lc00245aCrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Sackmann EK, Berthier E, Young EW et al (2012) Microfluidic kit-on-a-lid: a versatile platform for neutrophil chemotaxis assays. Blood 120(14):e45–e53.  https://doi.org/10.1182/blood-2012-03-416453CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Jones CN, Dalli J, Dimisko L et al (2012) Microfluidic chambers for monitoring leukocyte trafficking and humanized nano-proresolving medicines interactions. Proc Natl Acad Sci U S A 109(50):20560–20565.  https://doi.org/10.1073/pnas.1210269109CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Schwarz J, Bierbaum V, Merrin J et al (2016) A microfluidic device for measuring cell migration towards substrate-bound and soluble chemokine gradients. Sci Rep 6:36440.  https://doi.org/10.1038/srep36440CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Jain NG, Wong EA, Aranyosi AJ et al (2015) Microfluidic mazes to characterize T-cell exploration patterns following activation in vitro. Integr Biol (Camb) 7(11):1423–1431.  https://doi.org/10.1039/c5ib00146cCrossRefGoogle Scholar
  9. 9.
    Chen YC, Allen SG, Ingram PN et al (2015) Single-cell migration chip for chemotaxis-based microfluidic selection of heterogeneous cell populations. Sci Rep 5:9980.  https://doi.org/10.1038/srep09980CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Wong IY, Javaid S, Wong EA et al (2014) Collective and individual migration following the epithelial-mesenchymal transition. Nat Mater 13(11):1063–1071.  https://doi.org/10.1038/nmat4062CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Boyden S (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466CrossRefGoogle Scholar
  12. 12.
    Jones CN, Hoang AN, Dimisko L et al (2014) Microfluidic platform for measuring neutrophil chemotaxis from unprocessed whole blood. J Vis Exp 88.  https://doi.org/10.3791/51215
  13. 13.
    Boneschansker L, Yan J, Wong E et al (2014) Microfluidic platform for the quantitative analysis of leukocyte migration signatures. Nat Commun 5:4787.  https://doi.org/10.1038/ncomms5787CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Ramos CD, Canetti C, Souto JT et al (2005) MIP-1alpha[CCL3] acting on the CCR1 receptor mediates neutrophil migration in immune inflammation via sequential release of TNF-alpha and LTB4. J Leukoc Biol 78(1):167–177.  https://doi.org/10.1189/jlb.0404237CrossRefPubMedGoogle Scholar
  15. 15.
    Taub DD, Conlon K, Lloyd AR et al (1993) Preferential migration of activated CD4+ and CD8+ T cells in response to MIP-1 alpha and MIP-1 beta. Science 260(5106):355–358CrossRefGoogle Scholar
  16. 16.
    Singer M, Sansonetti PJ (2004) IL-8 is a key chemokine regulating neutrophil recruitment in a new mouse model of Shigella-induced colitis. J Immunol 173(6):4197–4206CrossRefGoogle Scholar
  17. 17.
    Becker H, Gartner C (2000) Polymer microfabrication methods for microfluidic analytical applications. Electrophoresis 21(1):12–26.  https://doi.org/10.1002/(sici)1522-2683(20000101)21:1<12::aid-elps12>3.0.co;2-7CrossRefPubMedGoogle Scholar
  18. 18.
    Schindelin J, Rueden CT, Hiner MC et al (2015) The ImageJ ecosystem: an open platform for biomedical image analysis. Mol Reprod Dev 82(7–8):518–529.  https://doi.org/10.1002/mrd.22489CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682.  https://doi.org/10.1038/nmeth.2019CrossRefPubMedGoogle Scholar
  20. 20.
    Tinevez JY, Perry N, Schindelin J et al (2017) TrackMate: an open and extensible platform for single-particle tracking. Methods 115:80–90.  https://doi.org/10.1016/j.ymeth.2016.09.016CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Brittany P. Boribong
    • 1
  • Amina Rahimi
    • 2
  • Caroline N. Jones
    • 3
    Email author
  1. 1.Genetics, Bioinformatics, and Computational BiologyVirginia TechBlacksburgUSA
  2. 2.Department of BiochemistryVirginia TechBlacksburgUSA
  3. 3.Department of Biological SciencesVirginia TechBlacksburgUSA

Personalised recommendations