{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T11:36:44Z","timestamp":1791459404062,"version":"4.3.4"},"reference-count":37,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,3,10]],"date-time":"2022-03-10T00:00:00Z","timestamp":1646870400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Thailand Science Research and Innovation (TSRI)","award":["2021"],"award-info":[{"award-number":["2021"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Micromachines"],"abstract":"<jats:p>This study proposes a microfluidic device used for X-\/Y-sperm separation based on monoclonal antibody-conjugated magnetic beads, which become positively charged in the flow system. Y-sperms were selectively captured via a monoclonal antibody and transferred onto the microfluidic device and were discarded, so that X-sperms can be isolated and commercially exploited for fertilization demands of female cattle in dairy industry. Therefore, the research team used monoclonal antibody-conjugated magnetic beads to increase the force that causes the Y-sperm to be pulled out of the system, leaving only the X-sperm for further use. The experimental design was divided into the following: Model 1, the microfluid system for sorting positive magnetic beads, which yielded 100% separation; Model 2, the sorting of monoclonal antibody-conjugated magnetic beads in the fluid system, yielding 98.84% microcirculation; Model 3, the sorting of monoclonal antibody-conjugated magnetic beads with sperm in the microfluid system, yielding 80.12% microcirculation. Moreover, the fabrication microfluidic system had thin film electrodes created via UV lithography and MWCNTs electrode structure capable of erecting an electrode wall 1500 \u00b5m above the floor with a flow channel width of only 100 \u00b5m. The system was tested using a constant flow rate of 2 \u00b5L\/min and X-\/Y-sperm were separated using carbon nanotube electrodes at 2.5 V. The structure created with the use of vertical electrodes and monoclonal antibody-conjugated magnetic beads technique produced a higher effective rejection effect and was able to remove a large number of unwanted sperm from the system with 80.12% efficiency.<\/jats:p>","DOI":"10.3390\/mi13030426","type":"journal-article","created":{"date-parts":[[2022,3,10]],"date-time":"2022-03-10T20:19:10Z","timestamp":1646943550000},"page":"426","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Antibody-Conjugated Magnetic Beads for Sperm Sexing Using a Multi-Wall Carbon Nanotube Microfluidic Device"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6176-5574","authenticated-orcid":false,"given":"Chalinee","family":"Phiphattanaphiphop","sequence":"first","affiliation":[{"name":"Industrial Engineering Department, Faculty of Engineering, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6877-1598","authenticated-orcid":false,"given":"Komgrit","family":"Leksakul","sequence":"additional","affiliation":[{"name":"Industrial Engineering Department, Faculty of Engineering, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thananut","family":"Wanta","sequence":"additional","affiliation":[{"name":"Industrial Engineering Department, Faculty of Engineering, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trisadee","family":"Khamlor","sequence":"additional","affiliation":[{"name":"Animal and Aquatic Science Department, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rungrueang","family":"Phattanakun","sequence":"additional","affiliation":[{"name":"Synchrotron Light Research Institute, Nakhon Ratchasima 30000, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.theriogenology.2007.04.005","article-title":"Overview of sexing sperm","volume":"68","author":"Seidel","year":"2007","journal-title":"Theriogenology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1002\/mrd.1120190205","article-title":"Separation of human X and Y spermatozoa by free-flow electrophoresis","volume":"19","author":"Engelmann","year":"1998","journal-title":"Gamete Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1111\/j.1365-2605.1991.tb01102.x","article-title":"Zeta potential of human X- and Y-bearing sperm","volume":"14","author":"Ishijima","year":"1991","journal-title":"Int. J. Androl."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"276","DOI":"10.2183\/pjab.59.276","article-title":"Separation of human X- and Y-bearing sperm using free-flow electrophoresis","volume":"59","author":"Kaneko","year":"1983","journal-title":"Proc. Jpn. Acad. Ser. B"},{"key":"ref_5","unstructured":"Wannaluk, T., Surat, H., Marninphan, T., Supamit, M., Wiwat, P., and Korawan, S. (2020). Production of single-chain fragment variable (scFv) antibodies specific to plasma membrane epitopes on bull Y-bearing sperm. Anim. Biotechnol., 1\u201311."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1613","DOI":"10.1002\/vms3.540","article-title":"Bovine semen sexing: Sperm embrane proteomics as candidates for immunological selection of X- and Ychromosome-bearing sperm","volume":"7","author":"Quelhas","year":"2021","journal-title":"Vet. Med. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1136\/vr.122.14.322","article-title":"Sexing of sperm by flow cytometry","volume":"122","author":"Morrell","year":"1988","journal-title":"Vet. Rec."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"375","DOI":"10.4141\/A03-022","article-title":"Past, present and future perspectives on sexing sperm","volume":"83","author":"Garner","year":"2003","journal-title":"Can. J. Anim. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e12821","DOI":"10.1111\/and.12821","article-title":"Selection of viable human spermatozoa with low levels of DNA fragmentation from an immotile population using density gradient centrifugation and magnetic-activated cell sorting","volume":"50","author":"Zhang","year":"2018","journal-title":"Andrologia"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"S336","DOI":"10.21037\/tau.2018.05.08","article-title":"Microfluidic-Based sperm sorting & analysis for treatment of male infertility","volume":"7","author":"Samuel","year":"2018","journal-title":"Transl. Androl. Urol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1063\/1.1685647","article-title":"Fluorescence activated cell sorting","volume":"43","author":"Bonner","year":"1972","journal-title":"Rev. Sci. Instrum."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1733","DOI":"10.1093\/oxfordjournals.humrep.a137925","article-title":"Gender preselection in humans Flow cytometric separation of X and Y spermatozoa for the prevention of X-linked diseases","volume":"8","author":"Johnson","year":"1993","journal-title":"Hum. Reprod."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1007\/s10544-007-9130-y","article-title":"DC-dielectrophoretic separation of biological cells by size","volume":"10","author":"Kang","year":"2008","journal-title":"Biomed. Microdevices"},{"key":"ref_14","unstructured":"Fleming, S., and Aitken, R.J. (2011). The Physical, Insemination, and Reproductive Quality of Honey Bee Queens (Apis mellifera L.), Springer."},{"key":"ref_15","unstructured":"Solomon, D.H., Caulfield, M.J., and Purss, H.K. (1986). Polymerization Process and Polymers Produced Thereby. (US4581429A), U.S. Patent."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1109\/JMEMS.2020.3020130","article-title":"Multiwalled carbon nanotubes in microfluidic chip for the separation of X- and Y-sperm based on a photolithography technique","volume":"29","author":"Phiphatanaphiphop","year":"2020","journal-title":"J. Microelectromech. Syst."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1016\/j.biotechadv.2016.01.007","article-title":"Sperm processing for advanced reproductive technologies: Where are we today?","volume":"34","author":"Rappa","year":"2016","journal-title":"Biotechnol. Adv."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1093\/humrep\/dem015","article-title":"Molecular glass wool filtration as a new tool for sperm preparation","volume":"22","author":"Grunewald","year":"2007","journal-title":"Hum. Reprod."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"134","DOI":"10.2164\/jandrol.107.003632","article-title":"Utility of magnetic cell separation as a molecular sperm preparation technique","volume":"29","author":"Said","year":"2008","journal-title":"J. Androl."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"17143","DOI":"10.1038\/s41598-020-73841-3","article-title":"A novel microfluidic chip-based sperm-sorting device constructed using design of experiment method","volume":"10","author":"Phiphattanaphiphop","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2535","DOI":"10.1039\/c1lc20236g","article-title":"Lensless imaging for simultaneous microfluidic sperm monitoring and sorting","volume":"11","author":"Zhang","year":"2011","journal-title":"Lab Chip"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Xiao, R.R., Yin, T., Zou, W., Tang, Y., Ding, J., and Yang, J. (2015). Generation of gradients on a microfluidic device: Toward a high-throughput investigation of spermatozoa chemotaxis. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0142555"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.anireprosci.2017.04.011","article-title":"Sperm sexing in Nili-Ravi buffalo through modified swim up: Validation using SYBR\u00ae green real-time PCR","volume":"182","author":"Awan","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1016\/S0015-0282(16)55681-7","article-title":"Sex preselection through albumin separation of sperm","volume":"59","author":"Beernink","year":"1993","journal-title":"Fertil. Steril."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1038\/246421a0","article-title":"Isolation of fractions rich in human Y sperm","volume":"246","author":"Ericsson","year":"1973","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.jevs.2018.04.005","article-title":"Sperm Sexing Mediated by Magnetic Nanoparticles in Donkeys, a Preliminary In Vitro Study","volume":"65","year":"2018","journal-title":"J. Equine Vet. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1292\/jvms.12-0303","article-title":"Application of a new convenience gender sorting method for mouse spermatozoa to mouse reproductive engineering technology","volume":"75","author":"Hashimoto","year":"2013","journal-title":"J. Vet. Med. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3552","DOI":"10.1002\/elps.200500298","article-title":"Focused electrophoretic motion and selected electrokinetic dispensing of particles and cells in cross-microchannels","volume":"26","author":"Xuan","year":"2005","journal-title":"Electrophoresis"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"18892","DOI":"10.1073\/pnas.0704958104","article-title":"Continuous inertial focusing, ordering, and separation of particles in microchannels","volume":"104","author":"Irimia","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1007\/s10404-006-0142-3","article-title":"Development of sorting, aligning, and orienting motile sperm using microfluidic device operated by hydrostatic pressure","volume":"3","author":"Seo","year":"2007","journal-title":"Microfluid. Nanofluid."},{"key":"ref_31","unstructured":"World Health Organization (2010). WHO Laboratory Manual for the Examination and Processing of Human Semenl."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1093\/oxfordjournals.humrep.a138198","article-title":"Centrifugation of human spermatozoa induces sublethal damage; separation of human spermatozoa from seminal plasma by a dextran swim-up procedure without centrifugation extends their motile lifetime","volume":"8","author":"Alvarez","year":"1993","journal-title":"Hum. Reprod."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1338","DOI":"10.1093\/humrep\/15.6.1338","article-title":"Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa","volume":"15","author":"Barroso","year":"2000","journal-title":"Hum. Reprod."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1002\/j.1939-4640.1988.tb01067.x","article-title":"Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques","volume":"9","author":"Aitken","year":"1988","journal-title":"J. Androl."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1093\/humrep\/13.5.1240","article-title":"The effects of antioxidant supplementation during percoll preparation on human sperm DNA integrity","volume":"13","author":"Hughes","year":"1998","journal-title":"Hum. Reprod."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1700197","DOI":"10.1002\/adbi.201700197","article-title":"Advances in microfluidics-based assisted reproductive technology: From sperm sorter to reproductive system-on-a-chip","volume":"2","author":"Kashaninejad","year":"2018","journal-title":"Adv. Biosyst."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/s41378-019-0072-3","article-title":"Rapid two-dimensional characterisation of proteins in solution","volume":"5","author":"Saar","year":"2019","journal-title":"Microsyst. Nanoeng."}],"container-title":["Micromachines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-666X\/13\/3\/426\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:34:13Z","timestamp":1760135653000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-666X\/13\/3\/426"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,10]]},"references-count":37,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["mi13030426"],"URL":"https:\/\/doi.org\/10.3390\/mi13030426","relation":{},"ISSN":["2072-666X"],"issn-type":[{"value":"2072-666X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,10]]}}}