The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results.

College of Agriculture and Life Sciences Nort Carolina State University Raleigh NC 27695 USA

Department of Gynecology and Obstetrics Faculty of Medicine University Hospital Brno Masaryk University Brno Jihlavská 20 625 00 Brno Czech Republic

Department of Human Morphology and Embryology Wroclaw Medical University Chałubińskiego 6a Str 50 368 Wroclaw Poland

Department of Veterinary Medicine Nicolaus Copernicus University Gagarina 7 Str 87 100 Toruń Poland

Laboratory of Reproductive Physiology Institute of Animal Biochemistry and Genetics Centre of Biosciences Slovak Academy of Sciences Dúbravská Cesta 9 840 05 Bratislava Slovakia

Veterinary Research Institute Hudcova 70 621 00 Brno Czech Republic

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Vaughan D.A., Leung A., Resetkova N., Ruthazer R., Penzias A.S., Sakkas D., Alper M.M. How many oocytes are optimal to achieve multiple live births with one stimulation cycle? The one-and-done approach. Fertil. Steril. 2017;107:397–404.e3. doi: 10.1016/j.fertnstert.2016.10.037. PubMed DOI

Giojalas L.C., Guidobaldi H.A. Getting to and away from the egg, an interplay between several sperm transport mechanisms and a complex oviduct physiology. Mol. Cell. Endocrinol. 2020;518:110954. doi: 10.1016/j.mce.2020.110954. PubMed DOI

Williams M., Thompson L.A., Li T.C., Mackenna A., Barratt C.L., Cooke I.D. Uterine flushing: A method to recover spermatozoa and leukocytes. Hum. Reprod. 1993;8:925–928. doi: 10.1093/oxfordjournals.humrep.a138168. PubMed DOI

Mortimer D., Leslie E.E., Kelly R.W., Templeton A.A. Morphological selection of human spermatozoa in vivo and in vitro. J. Reprod. Fertil. 1982;64:391–399. doi: 10.1530/jrf.0.0640391. PubMed DOI

Pérez-Cerezales S., Ramos-Ibeas P., Acuña O.S., Avilés M., Coy P., Rizos D., Gutiérrez-Adán A. The oviduct: From sperm selection to the epigenetic landscape of the embryo. Biol. Reprod. 2018;98:262–276. doi: 10.1093/biolre/iox173. PubMed DOI

Sakkas D., Ramalingam M., Garrido N., Barratt C.L. Sperm selection in natural conception: What can we learn from mother nature to improve assisted reproduction outcomes? Hum. Reprod. Update. 2015;21:711–726. doi: 10.1093/humupd/dmv042. PubMed DOI PMC

Baldini D., Ferri D., Baldini G.M., Lot D., Catino A., Vizziello D., Vizziello G. Sperm selection for ICSI: Do we have a winner? Cells. 2021;10:3566. doi: 10.3390/cells10123566. PubMed DOI PMC

Henkel R. Sperm preparation: State-of-the-art-physiological aspects and application of advanced sperm preparation methods. Asian J. Androl. 2012;14:260–269. doi: 10.1038/aja.2011.133. PubMed DOI PMC

Dai C., Zhang Z., Shan G., Chu L.T., Huang Z., Moskovtsev S., Librach C., Jarvi K., Sun Y. Advances in sperm analysis: Techniques, discoveries and applications. Nat. Rev. Urol. 2021;18:447–467. doi: 10.1038/s41585-021-00472-2. PubMed DOI

Dehghanpour F., Khalili M.A., Mangoli E., Talebi A.R., Anbari F., Shamsi F., Woodward B., Doostabadi M.R. Free centrifuge sorting method for high-count sperm preparation improves biological characteristics of human spermatozoa and clinical outcome: A sibling oocytes study. Andrologia. 2022;54:e14554. doi: 10.1111/and.14554. PubMed DOI

Zini A., Finelli A., Phang D., Jarvi K. Influence of semen processing technique on human sperm DNA integrity. Urology. 2000;56:1081–1084. doi: 10.1016/S0090-4295(00)00770-6. PubMed DOI

Yamanaka M., Tomita K., Hashimoto S., Matsumoto H., Satoh M., Kato H., Hosoi Y., Inoue M., Nakaoka Y., Morimoto Y. Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms. J. Reprod. Dev. 2016;62:599–606. doi: 10.1262/jrd.2016-112. PubMed DOI PMC

Gandini L., Lombardo F., Paoli D., Caruso F., Eleuteri P., Leter G., Ciriminna R., Culasso F., Dondero F., Lenzi A., et al. Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage. Hum. Reprod. 2004;19:1409–1417. doi: 10.1093/humrep/deh233. PubMed DOI

Sakkas D., Alvarez J.G. Sperm DNA fragmentation: Mechanisms of origin, impact on reproductive outcome, and analysis. Fertil. Steril. 2010;93:1027–1036. doi: 10.1016/j.fertnstert.2009.10.046. PubMed DOI

Gotsiridze K., Nana M., Mariam M., Tamar J. Live motile sperm sorting device improves embryo aneuploidy: A Retrospective Cohort Study. Fertil. Reprod. 2024;06:117–122. doi: 10.1142/S2661318224500166. DOI

Quinn M.M., Jalalian L., Ribeiro S., Ona K., Demirci U., Cedars M.I., Rosen M.P. Microfluidic sorting selects sperm for clinical use with reduced DNA damage compared to density gradient centrifugation with swim-up in split semen samples. Hum. Reprod. 2018;33:1388–1393. doi: 10.1093/humrep/dey239. PubMed DOI

Sheibak N., Amjadi F., Shamloo A., Zarei F., Zandieh Z. Microfluidic sperm sorting selects a subpopulation of high-quality sperm with a higher potential for fertilization. Hum. Reprod. 2024;39:902–911. doi: 10.1093/humrep/deae045. PubMed DOI

Banti M., Van Zyl E., Kafetzis D. Suprem preparation with microfluidic sperm sorting chip may improve intracytoplasmic sperm injection outcomes compared to density gradient centrifugation. Reprod. Sci. 2024;31:1695–1704. doi: 10.1007/s43032-024-01483-1. PubMed DOI PMC

Anbari F., Khalili M.A., Sultan Ahamed A.M., Mangoli E., Nabi A., Dehghanpour F., Sabour M. Microfluidic sperm selection yields higher sperm quality compared to conventional method in ICSI program: A pilot study. Syst. Biol. Reprod. Med. 2021;67:137–143. doi: 10.1080/19396368.2020.1837994. PubMed DOI

Hsu C.T., Lee C.I., Lin F.S., Wang F.Z., Chang H.C., Wang T.E., Huang C.C., Tsao H.M., Lee M.S., Agarwal A. Live motile sperm sorting device for enhanced sperm-fertilization competency: Comparative analysis with density-gradient centrifugation and microfluidic sperm sorting. J. Assist. Reprod. Genet. 2023;40:1855–1864. doi: 10.1007/s10815-023-02838-4. PubMed DOI PMC

Feyzioglu B.S., Avul Z. Effects of sperm separation methods before intrauterine insemination on pregnancy outcomes and live birth rates: Differences between the swim-up and microfluidic chip techniques. Medicine. 2023;102:e36042. doi: 10.1097/MD.0000000000036042. PubMed DOI PMC

Leisinger C.A., Adaniya G., Freeman M.R., Behnke E.J., Aguirre M., VerMilyea M.D., Schiewe M.C. Effect of microfluidic sperm separation vs. standard sperm washing processes on laboratory outcomes and clinical pregnancy rates in an unselected patient population. Reprod. Med. 2021;2:125–130. doi: 10.3390/reprodmed2030013. DOI

Pujianto D.A., Oktarina M., Sharaswati I.A. Hydrogen peroxide has adverse effects on human sperm quality parameters, induces apoptosis, and reduces survival. J. Hum. Reprod. Sci. 2021;14:121–128. doi: 10.4103/jhrs.jhrs_241_20. PubMed DOI PMC

Andrabi S.W., Ara A., Saharan A., Jaffar M., Gugnani N., Esteves S.C. Sperm DNA fragmentation: Causes, evaluation and management in male infertility. JBRA Assist. Reprod. 2024;28:306–319. doi: 10.5935/1518-0557.20230076. PubMed DOI PMC

Wright C., Milne S., Leeson H. Sperm DNA damage caused by oxidative stress: Modifiable clinical, lifestyle and nutritional factors in male infertility. Reprod. Biomed. Online. 2014;28:684–703. doi: 10.1016/j.rbmo.2014.02.004. PubMed DOI

Vasilescu S.A., Ding L., Parast F.Y., Nosrati R., Warkiani M.E. Sperm quality metrics were improved by a biomimetic microfluidic selection platform compared to swim-up methods. Microsyst. Nanoeng. 2023;9:37. doi: 10.1038/s41378-023-00501-7. PubMed DOI PMC

Kocur O.M., Xie P., Cheung S., Souness S., McKnight M., Rosenwaks Z., Palermo G.D. Can a sperm selection technique improve embryo ploidy? Andrology. 2023;11:1605–1612. doi: 10.1111/andr.13362. PubMed DOI PMC

Parrella A., Keating D., Cheung S., Xie P., Stewart J.D., Rosenwaks Z., Palermo G.D. A treatment approach for couples with disrupted sperm DNA integrity and recurrent ART failure. J. Assist. Reprod. Genet. 2019;36:2057–2066. doi: 10.1007/s10815-019-01543-5. PubMed DOI PMC

Shirota K., Yotsumoto F., Itoh H., Obama H., Hidaka N., Nakajima K., Miyamoto S. Separation efficiency of a microfluidic sperm sorter to minimize sperm DNA damage. Fertil. Steril. 2016;105:315–321.e1. doi: 10.1016/j.fertnstert.2015.10.023. PubMed DOI

Muratori M., Tarozzi N., Cambi M., Boni L., Iorio A.L., Passaro C., Luppino B., Nadalini M., Marchiani S., Tamburrino L., et al. Variation of DNA fragmentation levels during density gradient sperm selection for assisted reproduction techniques: A possible new male predictive parameter of pregnancy? Medicine. 2016;95:e3624. doi: 10.1097/MD.0000000000003624. PubMed DOI PMC

Takeshima T., Yumura Y., Kuroda S., Kawahara T., Uemura H., Iwasaki A. Effect of density gradient centrifugation on reactive oxygen species in human semen. Syst. Biol. Reprod. Med. 2017;63:192–198. doi: 10.1080/19396368.2017.1294214. PubMed DOI

Raad G., Bakos H.W., Bazzi M., Mourad Y., Fakih F., Shayya S., Mchantaf L., Fakih C. Differential impact of four sperm preparation techniques on sperm motility, morphology, DNA fragmentation, acrosome status, oxidative stress, and mitochondrial activity: A prospective study. Andrology. 2021;9:1549–1559. doi: 10.1111/andr.13038. PubMed DOI

Esteves S.C., Sharma R.K., Thomas A.J., Agarwal A. Effect of swim-up sperm washing and subsequent capacitation on acrosome status and functional membrane integrity of normal sperm. Int. J. Fertil. Womens Med. 2000;45:335–341. PubMed

Paoli D., Gallo M., Rizzo F., Baldi E., Francavilla S., Lenzi A., Lombardo F., Gandini L. Mitochondrial membrane potential profile and its correlation with increasing sperm motility. Fertil. Steril. 2011;95:2315–2319. doi: 10.1016/j.fertnstert.2011.03.059. PubMed DOI

Agarwal A., Saleh R.A. Role of oxidants in male infertility: Rationale, significance, and treatment. Urol. Clin. N. Am. 2002;29:817–827. doi: 10.1016/S0094-0143(02)00081-2. PubMed DOI

Ferreira Aderaldo J., da Silva Maranhão K., Ferreira Lanza D.C. Does microfluidic sperm selection improve clinical pregnancy and miscarriage outcomes in assisted reproductive treatments? A systematic review and meta-analysis. PLoS ONE. 2023;18:e0292891. doi: 10.1371/journal.pone.0292891. PubMed DOI PMC

Jahangiri A.R., Ziarati N., Dadkhah E., Bucak M.N., Rahimizadeh P., Shahverdi A., Sadighi Gilani M.A., Topraggaleh T.R. Microfluidics: The future of sperm selection in assisted reproduction. Andrology. 2024;12:1236–1252. doi: 10.1111/andr.13578. PubMed DOI

World Health Organization . WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. WHO Press; Geneva, Switzerland: 2010. pp. 1–271.

Doostabadi M.R., Mangoli E., Marvast L.D., Dehghanpour F., Maleki B., Torkashvand H., Talebi A.R. Microfluidic devices employing chemo- and thermotaxis for sperm selection can improve sperm parameters and function in patients with high DNA fragmentation. Andrologia. 2022;54:e14623. doi: 10.1111/and.14623. PubMed DOI

Estevez S.C., Verza S. Relationship of in vitro acrosome reacton to sperm function: An update. Open Rep. Sci. J. 2011;3:72–84. doi: 10.2174/1874255601103010072. DOI

Gangwar C., Kharche S.D., Mishra A.K., Saraswat S., Kumar N., Sikarwar A.K. Effect of diluent sugars on capacitation status and acrosome reaction of spermatozoa in buck semen at refrigerated temperature. Trop. Anim. Health Prod. 2020;52:3409–3415. doi: 10.1007/s11250-020-02374-8. PubMed DOI

Smiley S.T., Reers M., Mottola-Hartshorn C., Lin M., Chen A., Smith T.W., Steele G.D., Chen L.B. Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. Proc. Natl. Acad. Sci. USA. 1991;88:3671–3675. doi: 10.1073/pnas.88.9.3671. PubMed DOI PMC