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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Microfabrication of low-cost customisable counting chambers for standardised estimation of sperm concentration

Yue Liu A B , Megan Chesnut B , Amy Guitreau B , Jacob Beckham A , Adam Melvin C , Jason Eades A , Terrence R. Tiersch B and William Todd Monroe https://orcid.org/0000-0002-7889-3799 A D
+ Author Affiliations
- Author Affiliations

A Department of Biological and Agricultural Engineering, Louisiana State University, E. B. Doran Building, Baton Rouge, LA 70803, USA.

B Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, 2288 Gourrier Avenue, Baton Rouge, LA 70820, USA.

C Cain Department of Chemical Engineering, Louisiana State University, 3314F Patrick F. Taylor Hall, Baton Rouge, LA 70803, USA.

D Corresponding author. Email: tmonroe@lsu.edu

Reproduction, Fertility and Development 32(9) 873-878 https://doi.org/10.1071/RD19154
Submitted: 3 May 2019  Accepted: 20 December 2019   Published: 27 May 2020

Abstract

Evaluation of sperm concentration is essential for research and procedures involving AI, cryopreservation and sperm quality assessment. Microfabrication technologies have shown tremendous potential for rapid prototyping and fabrication of devices to assist reproduction and fertility research, but such utility has not yet been made available for most reproduction laboratories. The aim of this study was to evaluate the feasibility of using microfabrication techniques to produce counting chambers for estimation of sperm concentration. Zebrafish (Danio rerio) spermatozoa were used as a model for evaluation of functionality of the chambers. These microfabricated enumeration grid chambers (MEGC) were composed of a polydimethylsiloxane (PDMS) coverslip with grid patterns (100 μm × 100 μm) and a PDMS base platform to create a known volume with a 10-μm height to restrict the cells to a single layer. The results of cell counts estimated by two of three prototype MEGC devices tested were not significantly different from the control device, a commercially available Makler chamber. The material cost for a MEGC was less than US$0.10 compared with product costs of approximately US$100 for a standard haemocytometer and US$700 for a Makler counting chamber. This study demonstrates the feasibility of microfabrication in creating low-cost counting chambers to enhance standardisation and strengthen interdisciplinary collaborations.

Graphical Abstract Image

Additional keywords: aquatic models, cryopreservation, zebrafish.


References

Beckham, J., Alam, F., Omojola, V., Scherr, T., Guitreau, A., Melvin, A., Park, D. S., Choi, J.-W., Tiersch, T. R., and Monroe, W. T. (2018). A microfluidic device for motility and osmolality analysis of zebrafish sperm. Biomed. Microdevices 20, 67.
A microfluidic device for motility and osmolality analysis of zebrafish sperm.Crossref | GoogleScholarGoogle Scholar | 30090952PubMed |

Christensen, P., Stryhn, H., and Hansen, C. (2005). Discrepancies in the determination of sperm concentration using Bürker-Türk, Thoma and Makler counting chambers. Theriogenology 63, 992–1003.
Discrepancies in the determination of sperm concentration using Bürker-Türk, Thoma and Makler counting chambers.Crossref | GoogleScholarGoogle Scholar | 15710187PubMed |

Hagedorn, M., Varga, Z., Walter, R. B., and Tiersch, T. R. (2019). Workshop report: cryopreservation of aquatic biomedical models. Cryobiology 86, 120–129.
Workshop report: cryopreservation of aquatic biomedical models.Crossref | GoogleScholarGoogle Scholar | 30389588PubMed |

Kjelland, M. E., Stroud, T., Ayliffe, H. E., and Dittami, G. (2014). 178 Portable automated microfluidic device for rapid determination of sperm counts. Reprod. Fertil. Dev. 26, 203–204.
178 Portable automated microfluidic device for rapid determination of sperm counts.Crossref | GoogleScholarGoogle Scholar |

Liu, Y., Torres, L., and Tiersch, T. R. (2018). Quality evaluation of sperm from livebearing fishes: standardized assessment of sperm bundles (spermatozeugmata) from Xenotoca eiseni (Goodeidae). Theriogenology 107, 50–56.
Quality evaluation of sperm from livebearing fishes: standardized assessment of sperm bundles (spermatozeugmata) from Xenotoca eiseni (Goodeidae).Crossref | GoogleScholarGoogle Scholar | 29128701PubMed |

Park, D. S., Egnatchik, R. A., Bordelon, H., Tiersch, T. R., and Monroe, W. T. (2012). Microfluidic mixing for sperm activation and motility analysis of pearl Danio zebrafish. Theriogenology 78, 334–344.
Microfluidic mixing for sperm activation and motility analysis of pearl Danio zebrafish.Crossref | GoogleScholarGoogle Scholar | 22494680PubMed |

Seaman, E. K., Goluboff, E., BarChama, N., and Fisch, H. (1996). Accuracy of semen counting chambers as determined by the use of latex beads. Fertil. Steril. 66, 662–665.
Accuracy of semen counting chambers as determined by the use of latex beads.Crossref | GoogleScholarGoogle Scholar | 8816636PubMed |

Sia, S. K., and Whitesides, G. M. (2003). Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies. Electrophoresis 24, 3563–3576.
Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies.Crossref | GoogleScholarGoogle Scholar | 14613181PubMed |

World Health Organization (WHO) (2010) ‘WHO Laboratory Manual for the Examination and Processing of Human Semen.’ 5th edn. (WHO Press: Geneva.)