Biomedical Microdevices

, 20:67 | Cite as

A microfluidic device for motility and osmolality analysis of zebrafish sperm

  • Jacob Beckham
  • Faiz Alam
  • Victor Omojola
  • Thomas Scherr
  • Amy Guitreau
  • Adam Melvin
  • Daniel S. Park
  • Jin-Woo Choi
  • Terrence R. Tiersch
  • W. Todd Monroe


A microfluidic chip is described that facilitates research and quality control analysis of zebrafish sperm which, due to its miniscule (i.e., 2–5 μl) sample volume and short duration of motility (i.e., <1 min), present a challenge for traditional manual assessment methods. A micromixer molded in polydimethylsiloxane (PDMS) bonded to a glass substrate was used to activate sperm samples by mixing with water, initiated by the user depressing a transfer pipette connected to the chip. Sample flow in the microfluidic viewing chamber was able to be halted within 1 s, allowing for rapid analysis of the sample using established computer-assisted sperm analysis (CASA) methods. Zebrafish sperm cell activation was consistent with manual hand mixing and yielded higher values of motility at earlier time points, as well as more subtle time-dependent trends in motility, than those processed by hand. Sperm activation curves, which indicate sample quality by evaluating percentage and duration of motility at various solution osmolalities, were generated with on-chip microfabricated gold floor electrodes interrogated by impedance spectroscopy. The magnitude of admittance was linearly proportional to osmolality and was not affected by the presence of sperm cells in the vicinity of the electrodes. This device represents a pivotal step in streamlining methods for consistent, rapid assessment of sperm quality for aquatic species. The capability to rapidly activate sperm and consistently measure motility with CASA using the microfluidic device described herein will help improve the reproducibility of studies on sperm and assist development of germplasm repositories.


Zebrafish Sperm Motility Impedance 



We thank L. Torres for assistance with sperm sample preparation, R.L. McCarley for access to equipment, and D.J. Hayes for insightful discussions. We also acknowledge support from the National Institutes of Health grants R24-RR023998, R24-OD011120 and 2R24-OD010441, and the National Institute of Food and Agriculture, United States Department of Agriculture (Hatch project LAB94231 and NC1194). This report was approved for publication by the Director of the Louisiana Agricultural Experiment Station as number 2017-241-31421.

Supplementary material

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  1. D.J. Beebe et al., Annu. Rev. Biomed. Eng. 4, 261–286 (2002). CrossRefGoogle Scholar
  2. S. Begolo et al., Lab Chip (2014). CrossRefGoogle Scholar
  3. Bruus, H., in Microscale Acoustofluidics, ed. By T. L. Laurell, Andreas (The Royal Society of Chemistry, 2015), 1-28 Google Scholar
  4. Y.-A. Chen et al., Microfluid. Nanofluid. 10, 59–67 (2010). CrossRefGoogle Scholar
  5. C.Y. Chen et al., Analyst 138, 4967–4974 (2013). CrossRefGoogle Scholar
  6. B.S. Cho et al., Anal. Chem. 75, 1671–1675 (2003). CrossRefGoogle Scholar
  7. J. Cosson, Aquac. Int. 12, 69–85 (2004)CrossRefGoogle Scholar
  8. K. Dooley, L.I. Zon, Curr. Opin. Genet. Dev. 10, 252–256 (2000). CrossRefGoogle Scholar
  9. M.D. Dryden, A.R. Wheeler, PLoS One 10, e0140349 (2015). CrossRefGoogle Scholar
  10. M.T. Glynn et al., Lab Chip 14, 2844–2851 (2014). CrossRefGoogle Scholar
  11. M. Hagedorn et al., Cryobiology 58, 12–19 (2009). CrossRefGoogle Scholar
  12. H. A. Herman, Improving cattle by the millions. NAAB and the development and worldwide application of artificial insemination, 1st Edition, (University of Missouri Press, Columbia, 1981)Google Scholar
  13. K. Iwai et al., Lab Chip 14, 3790–3799 (2014). CrossRefGoogle Scholar
  14. W. Li et al., Lab Chip 12, 1587–1590 (2012). CrossRefGoogle Scholar
  15. G.J. Lieschke, P.D. Currie, Nat. Rev. Genet. 8, 353–367 (2007). CrossRefGoogle Scholar
  16. MicroChem, Permanent Epoxy Negative Photoresist Processing Guidelines for SU-2025. (MicroChem, 2011), Accessed May 12, 2013 
  17. National Institutes of Health, Research Portfolio Online Reporting Tools. (National Institutes of Health, Bethesda, MD, 2018), Accessed May 5, 2018
  18. C.E. Nwankire et al., Biosens. Bioelectron. 68, 382–389 (2015). CrossRefGoogle Scholar
  19. D.S. Park et al., Theriogenology 78, 334–344 (2012). CrossRefGoogle Scholar
  20. A.A. Rowe et al., PLoS One 6, e23783 (2011). CrossRefGoogle Scholar
  21. I.F. Sbalzarini, P. Koumoutsakos, J. Struct. Biol. 151, 182–195 (2005). CrossRefGoogle Scholar
  22. T. Scherr et al., J. Micromech. Microeng. 22, 55019 (2012). CrossRefGoogle Scholar
  23. T. Scherr et al., Biomed. Microdevices 17, 65 (2015). CrossRefGoogle Scholar
  24. J. Schindelin et al., Nat. Methods 9, 676–682 (2012). CrossRefGoogle Scholar
  25. C.A. Schneider et al., Nat. Methods 9, 671–675 (2012)CrossRefGoogle Scholar
  26. C. Seguin et al., Appl. Surf. Sci. 256, 2524–2531 (2010). CrossRefGoogle Scholar
  27. D.B. Seo et al., Microfluid. Nanofluid. 3, 561–570 (2007). CrossRefGoogle Scholar
  28. S. Sugiura et al., Anal. Chem. 82, 8278–8282 (2010). CrossRefGoogle Scholar
  29. R.S. Suh et al., Hum. Reprod. 21, 477–483 (2006). CrossRefGoogle Scholar
  30. G. Sui et al., Anal. Chem. 78, 5543–5551 (2006). CrossRefGoogle Scholar
  31. T.R. Tiersch, C.C. Green Cryopreservation in Aquatic Species, 2nd edition. World Aquaculture Society, Advances in World Aquaculture, Baton Rouge, Louisiana, 1003 pages (2011)Google Scholar
  32. T.R. Tiersch et al., Soc. Reprod. Fertil. Suppl. 65, 493–508 (2007)Google Scholar
  33. L. Torres, T.R. Tiersch, J. World Aquacult. Soc. (In Press) (2018). CrossRefGoogle Scholar
  34. C. Tropea et al., Springer handbook of experimental fluid mechanics, 1st Edition, (Springer Science & Business Media, Berlin, 2007)Google Scholar
  35. H. Wang et al., Lab Chip 17, 1264–1269 (2017). CrossRefGoogle Scholar
  36. J.G. Wilson-Leedy, R.L. Ingermann, Theriogenology 67, 661–672 (2007). CrossRefGoogle Scholar
  37. J.G. Wilson-Leedy et al., Theriogenology 71, 1054–1062 (2009). CrossRefGoogle Scholar
  38. J.S. Wolenski, N.H. Hart, J. Exp. Zool. 243, 259–273 (1987)CrossRefGoogle Scholar
  39. L. Xie et al., Clin. Chem. 56, 1270–1278 (2010). CrossRefGoogle Scholar
  40. H. Yang et al., Theriogenology 68, 128–136 (2007). CrossRefGoogle Scholar
  41. Zebrafish Information Network (ZFIN). (University of Oregon, Eugene, 2018), Accessed May 5, 2018Google Scholar
  42. Zebrafish International Resource Center, Dry Food Recipe for fish 90+ dpf. (University of Oregon, 2015), Accessed May 10, 2015

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biological & Agricultural EngineeringLouisiana State University and Agricultural CenterBaton RougeUSA
  2. 2.Department of ChemistryVanderbilt UniversityNashvilleUSA
  3. 3.Aquatic Germplasm and Genetic Resources CenterLouisiana State University Agricultural CenterBaton RougeUSA
  4. 4.Cain Department of Chemical EngineeringLouisiana State UniversityBaton RougeUSA
  5. 5.Department of Mechanical EngineeringLouisiana State UniversityBaton RougeUSA
  6. 6.School of Electrical Engineering & Computer ScienceLouisiana State UniversityBaton RougeUSA

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