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 tissues toward sites of infections along the chemokine gradients, in a process named chemotaxis. Studying neutrophil chemotaxis using conventional tools, such as a transwell assay, often requires isolation of neutrophils from whole blood. This process requires milliliters of blood, trained personnel, and can easily alter the ability of chemotaxis. Microfluidics is an enabling technology for studying chemotaxis of neutrophils in vitro with high temporal and spatial resolution. In this chapter, we describe a procedure for probing human neutrophil chemotaxis directly in one droplet of whole blood, without neutrophil isolation, using microfluidic devices. The same devices can be applied to the study the chemotaxis of neutrophils from small animals, e.g., mice and rats.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
de Oliveira S, Rosowski EE, Huttenlocher A (2016) Neutrophil migration in infection and wound repair: going forward in reverse. Nat Rev Immunol 16(6):378–391. https://doi.org/10.1038/nri.2016.49
Dinauer MC (2014) Disorders of neutrophil function: an overview. Methods Mol Biol 1124:501–515. https://doi.org/10.1007/978-1-62703-845-4_30
Butler KL, Ambravaneswaran V, Agrawal N, Bilodeau M, Toner M, Tompkins RG, Fagan S, Irimia D (2010) Burn injury reduces neutrophil directional migration speed in microfluidic devices. PLoS One 5(7):e11921. https://doi.org/10.1371/journal.pone.0011921
LeBlanc AK, LeBlanc CJ, Rohrbach BW, Kania SA (2015) Serial evaluation of neutrophil function in tumour-bearing dogs undergoing chemotherapy. Vet Comp Oncol 13(1):20–27. https://doi.org/10.1111/vco.12015
Soehnlein O, Steffens S, Hidalgo A, Weber C (2017) Neutrophils as protagonists and targets in chronic inflammation. Nat Rev Immunol 17(4):248–261. https://doi.org/10.1038/nri.2017.10
Brown KA, Brain SD, Pearson JD, Edgeworth JD, Lewis SM, Treacher DF (2006) Neutrophils in development of multiple organ failure in sepsis. Lancet 368(9530):157–169. https://doi.org/10.1016/S0140-6736(06)69005-3
Boyden S (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466
Zigmond SH (1977) Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors. J Cell Biol 75(2 Pt 1):606–616
Zicha D, Dunn GA, Brown AF (1991) A new direct-viewing chemotaxis chamber. J Cell Sci 99(Pt 4):769–775
Servant G, Weiner OD, Herzmark P, Balla T, Sedat JW, Bourne HR (2000) Polarization of chemoattractant receptor signaling during neutrophil chemotaxis. Science 287(5455):1037–1040
Whitesides GM (2006) The origins and the future of microfluidics. Nature 442(7101):368–373. https://doi.org/10.1038/nature05058
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.5RU0216-056R
Irimia D, Geba DA, Toner M (2006) Universal microfluidic gradient generator. Anal Chem 78(10):3472–3477. https://doi.org/10.1021/ac0518710
Irimia D, Charras G, Agrawal N, Mitchison T, Toner M (2007) Polar stimulation and constrained cell migration in microfluidic channels. Lab Chip 7(12):1783–1790. https://doi.org/10.1039/b710524j
Agrawal N, Toner M, Irimia D (2008) Neutrophil migration assay from a drop of blood. Lab Chip 8(12):2054–2061. https://doi.org/10.1039/b813588f
Hamza B, Wong E, Patel S, Cho H, Martel J, Irimia D (2014) Retrotaxis of human neutrophils during mechanical confinement inside microfluidic channels. Integrat Biol 6(2):175–183. https://doi.org/10.1039/c3ib40175h
Hoang AN, Jones CN, Dimisko L, Hamza B, Martel J, Kojic N, Irimia D (2013) Measuring neutrophil speed and directionality during chemotaxis, directly from a droplet of whole blood. Technology 1(1):49. https://doi.org/10.1142/S2339547813500040
Jones CN, Hoang AN, Dimisko L, Hamza B, Martel J, Irimia D (2014) Microfluidic platform for measuring neutrophil chemotaxis from unprocessed whole blood. J Vis Exp (88). https://doi.org/10.3791/51215
Kasuga K, Yang R, Porter TF, Agrawal N, Petasis NA, Irimia D, Toner M, Serhan CN (2008) Rapid appearance of resolvin precursors in inflammatory exudates: novel mechanisms in resolution. J Immunol 181(12):8677–8687
Sackmann EK, Berthier E, Schwantes EA, Fichtinger PS, Evans MD, Dziadzio LL, Huttenlocher A, Mathur SK, Beebe DJ (2014) Characterizing asthma from a drop of blood using neutrophil chemotaxis. Proc Natl Acad Sci U S A 111(16):5813–5818. https://doi.org/10.1073/pnas.1324043111
Sackmann EK, Berthier E, Young EW, Shelef MA, Wernimont SA, Huttenlocher A, Beebe DJ (2012) Microfluidic kit-on-a-lid: a versatile platform for neutrophil chemotaxis assays. Blood 120(14):e45–e53. https://doi.org/10.1182/blood-2012-03-416453
Jones CN, Hoang AN, Martel JM, Dimisko L, Mikkola A, Inoue Y, Kuriyama N, Yamada M, Hamza B, Kaneki M, Warren HS, Brown DE, Irimia D (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.5TA0715-310RR
Mankovich AR, Lee CY, Heinrich V (2013) Differential effects of serum heat treatment on chemotaxis and phagocytosis by human neutrophils. PLoS One 8(1):e54735. https://doi.org/10.1371/journal.pone.0054735
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Wang, X., Irimia, D. (2018). Neutrophil Chemotaxis in One Droplet of Blood Using Microfluidic Assays. In: Gautreau, A. (eds) Cell Migration. Methods in Molecular Biology, vol 1749. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7701-7_25
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7701-7_25
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7700-0
Online ISBN: 978-1-4939-7701-7
eBook Packages: Springer Protocols