{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T23:56:21Z","timestamp":1769644581231,"version":"3.49.0"},"reference-count":48,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2020,10,19]],"date-time":"2020-10-19T00:00:00Z","timestamp":1603065600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF-2018M3C7A1022309"],"award-info":[{"award-number":["NRF-2018M3C7A1022309"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Along with the increasing popularity of larval zebrafish as an experimental animal in the fields of drug screening, neuroscience, genetics, and developmental biology, the need for tools to deal with multiple larvae has emerged. Microfluidic channels have been employed to handle multiple larvae simultaneously, even for sensing electroencephalogram (EEG). In this study, we developed a microfluidic chip capable of uniform and continuous drug infusion across all microfluidic channels during EEG recording. Owing to the modular design of the microfluidic channels, the number of animals under investigation can be easily increased. Using the optimized design of the microfluidic chip, liquids could be exchanged uniformly across all channels without physically affecting the larvae contained in the channels, which assured a stable environment maintained all the time during EEG recording, by eliminating environmental artifacts and leaving only biological effects to be seen. To demonstrate the usefulness of the developed system in drug screening, we continuously measured EEG from four larvae without and with pentylenetetrazole application, up to 60 min. In addition, we recorded EEG from valproic acid (VPA)-treated zebrafish and demonstrated the suppression of seizure by VPA. The developed microfluidic system could contribute to the mass screening of EEG for drug development to treat neurological disorders such as epilepsy in a short time, owing to its handy size, cheap fabrication cost, and the guaranteed uniform drug infusion across all channels with no environmentally induced artifacts.<\/jats:p>","DOI":"10.3390\/s20205903","type":"journal-article","created":{"date-parts":[[2020,10,19]],"date-time":"2020-10-19T09:36:28Z","timestamp":1603100188000},"page":"5903","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["A Microfluidic System for Stable and Continuous EEG Monitoring from Multiple Larval Zebrafish"],"prefix":"10.3390","volume":"20","author":[{"given":"Yuhyun","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 333, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0646-9806","authenticated-orcid":false,"given":"Hee Won","family":"Seo","sequence":"additional","affiliation":[{"name":"Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 333, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2235-3312","authenticated-orcid":false,"given":"Kyeong Jae","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 333, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jae-Won","family":"Jang","sequence":"additional","affiliation":[{"name":"Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 333, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4926-8978","authenticated-orcid":false,"given":"Sohee","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 333, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bromberg, Y. (2013). Chapter 15: Disease Gene Prioritization. PLoS Comput. Biol., 9.","DOI":"10.1371\/journal.pcbi.1002902"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2065","DOI":"10.1152\/jn.00538.2015","article-title":"Zebrafish needle EMG: A new tool for high-throughput drug screens","volume":"114","author":"Cho","year":"2015","journal-title":"J. Neurophysiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1038\/nn.3934","article-title":"Animal models in epilepsy research: Legacies and new directions","volume":"18","author":"Grone","year":"2015","journal-title":"Nat. Neurosci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1038\/nature12111","article-title":"The zebrafish reference genome sequence and its relationship to the human genome","volume":"496","author":"Howe","year":"2013","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1039\/C6LC00044D","article-title":"Fish-on-a-chip: Microfluidics for zebrafish research","volume":"16","author":"Yang","year":"2016","journal-title":"Lab Chip"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.knosys.2013.02.014","article-title":"Automated EEG analysis of epilepsy: A review","volume":"45","author":"Acharya","year":"2013","journal-title":"Knowl. Based Syst."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3099","DOI":"10.1038\/s41598-017-03482-6","article-title":"Zebrafish as an animal model in epilepsy studies with multichannel EEG recordings","volume":"7","author":"Cho","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1016\/S0140-6736(14)60456-6","article-title":"Epilepsy: New advances","volume":"385","author":"Perucca","year":"2015","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.jneumeth.2011.06.001","article-title":"Recording the adult zebrafish cerebral field potential during pentylenetetrazole seizures","volume":"200","author":"Pineda","year":"2011","journal-title":"J. Neurosci. Methods"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Afrikanova, T., Serruys, A.S.K., Buenafe, O.E.M., Clinckers, R., Smolders, I., de Witte, P.A.M., Crawford, A.D., and Esguerra, C.V. (2013). Validation of the Zebrafish Pentylenetetrazol Seizure Model: Locomotor versus Electrographic Responses to Antiepileptic Drugs. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0054166"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1016\/j.neuroscience.2004.11.031","article-title":"Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression","volume":"131","author":"Baraban","year":"2005","journal-title":"Neuroscience"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1523\/ENEURO.0066-17.2017","article-title":"Behavioral Comorbidities and Drug Treatments in a Zebrafish scn1lab Model of Dravet Syndrome","volume":"4","author":"Grone","year":"2017","journal-title":"eNeuro"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.brainresbull.2011.11.020","article-title":"Perspectives of zebrafish models of epilepsy: What, how and where next?","volume":"87","author":"Stewart","year":"2012","journal-title":"Brain Res. Bull."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1111\/j.1528-1167.2007.01075.x","article-title":"A Large-scale Mutagenesis Screen to Identify Seizure-resistant Zebrafish","volume":"48","author":"Baraban","year":"2007","journal-title":"Epilepsia"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2410","DOI":"10.1038\/ncomms3410","article-title":"ARTICLE Drug screening in Scn1a zebrafish mutant identifies clemizole as a potential Dravet syndrome treatment","volume":"4","author":"Baraban","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1242\/dmm.002139","article-title":"Zebrafish as a model for studying genetic aspects of epilepsy","volume":"3","author":"Hortopan","year":"2010","journal-title":"DMM Dis. Model. Mech."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.expneurol.2012.06.013","article-title":"A novel zebrafish model of hyperthermia-induced seizures reveals a role for TRPV4 channels and NMDA-type glutamate receptors","volume":"237","author":"Hunt","year":"2012","journal-title":"Exp. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1093\/brain\/awu133","article-title":"On the nature of seizure dynamics","volume":"137","author":"Jirsa","year":"2014","journal-title":"Brain"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1732","DOI":"10.1039\/c3lc50099c","article-title":"Zebrafish Entrapment by Restriction Array (ZEBRA) device: A low-cost, agarose-free zebrafish mounting technique for automated imaging","volume":"13","author":"Bischel","year":"2013","journal-title":"Lab Chip"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Meyer, M., Dhamne, S.C., LaCoursiere, C.M., Tambunan, D., Poduri, A., and Rotenberg, A. (2016). Microarray Noninvasive Neuronal Seizure Recordings from Intact Larval Zebrafish. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0159472"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"28248","DOI":"10.1038\/srep28248","article-title":"A Novel Long-term, Multi-Channel and Non-invasive Electrophysiology Platform for Zebrafish","volume":"6","author":"Hong","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.neuroimage.2009.02.028","article-title":"Signal quality of simultaneously recorded invasive and non-invasive EEG","volume":"46","author":"Ball","year":"2009","journal-title":"Neuroimage"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1038\/s41467-017-02404-4","article-title":"Brain activity patterns in high-throughput electrophysiology screen predict both drug efficacies and side effects","volume":"9","author":"Eimon","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"127332","DOI":"10.1016\/j.snb.2019.127332","article-title":"High-throughput zebrafish intramuscular recording assay","volume":"304","author":"Cho","year":"2020","journal-title":"Sens. Actuators B Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1002\/biot.201200129","article-title":"Microfluidic tools for developmental studies of small model organisms--nematodes, fruit flies, and zebrafish","volume":"8","author":"Hwang","year":"2013","journal-title":"Biotechnol. J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"892","DOI":"10.1039\/C1LC20351G","article-title":"Fish and chips: A microfluidic perfusion platform for monitoring zebrafish development","volume":"12","author":"Choudhury","year":"2012","journal-title":"Lab Chip"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1016\/j.snb.2017.10.050","article-title":"Lab-on-a-Chip imaging micro-echocardiography (i\u03bcEC) for rapid assessment of cardiovascular activity in zebrafish larvae","volume":"256","author":"Fuad","year":"2018","journal-title":"Sens. Actuators B Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1039\/C4LC01186D","article-title":"High-throughput mapping of brain-wide activity in awake and drug-responsive vertebrates","volume":"15","author":"Lin","year":"2015","journal-title":"Lab Chip"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2017\/9053764","article-title":"A 3D-Printed Sensor for Monitoring Biosignals in Small Animals","volume":"2017","author":"Cho","year":"2017","journal-title":"J. Healthc. Eng."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"aww342","DOI":"10.1093\/brain\/aww342","article-title":"Clemizole and modulators of serotonin signalling suppress seizures in Dravet syndrome","volume":"140","author":"Griffin","year":"2017","journal-title":"Brain"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fphar.2018.00573","article-title":"Preclinical animal models for Dravet syndrome: Seizure phenotypes, comorbidities and drug screening","volume":"9","author":"Griffin","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Collymore, C., Rasmussen, S., and Tolwani, R.J. (2013). Gavaging adult zebrafish. J. Vis. Exp., 1\u20134.","DOI":"10.3791\/50691-v"},{"key":"ref_33","first-page":"198","article-title":"Efficacy and safety of 5 anesthetics in adult zebrafish (Danio rerio)","volume":"53","author":"Collymore","year":"2014","journal-title":"J. Am. Assoc. Lab. Anim. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1093\/ilar.53.2.192","article-title":"Anesthesia and euthanasia in zebrafish","volume":"53","author":"Matthews","year":"2012","journal-title":"ILAR J."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Johnston, L., Ball, R.E., Acuff, S., Gaudet, J., Sornborger, A., and Lauderdale, J.D. (2013). Electrophysiological recording in the brain of intact adult zebrafish. J. Vis. Exp., 1\u201311.","DOI":"10.3791\/51065"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.eplepsyres.2007.03.015","article-title":"Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants","volume":"75","author":"Berghmans","year":"2007","journal-title":"Epilepsy Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.ejphar.2010.06.041","article-title":"Improvement of pentylenetetrazol-induced learning deficits by valproic acid in the adult zebrafish","volume":"643","author":"Lee","year":"2010","journal-title":"Eur. J. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1007\/s13238-015-0206-9","article-title":"Drug screening: Zebrafish as a tool for studying epileptic-related chemical compounds","volume":"6","author":"Zhao","year":"2015","journal-title":"Protein Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"112315","DOI":"10.1016\/j.bios.2020.112315","article-title":"An EEG system to detect brain signals from multiple adult zebrafish","volume":"164","author":"Lee","year":"2020","journal-title":"Biosens. Bioelectron."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Aydin, \u00dc., Vorwerk, J., D\u00fcmpelmann, M., K\u00fcpper, P., Kugel, H., Heers, M., Wellmer, J., Kellinghaus, C., Haueisen, J., and Rampp, S. (2015). Combined EEG\/MEG can outperform single modality EEG or MEG source reconstruction in presurgical epilepsy diagnosis. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0118753"},{"key":"ref_41","first-page":"773","article-title":"Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures","volume":"5","author":"Baxendale","year":"2012","journal-title":"Dis. Model. Mech."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.neuint.2017.11.011","article-title":"Neurochemistry International Methylated fl avonoids as anti-seizure agents: Naringenin 4\u2032, 7-dimethyl ether attenuates epileptic seizures in zebra fish and mouse models","volume":"112","author":"Steurs","year":"2018","journal-title":"Neurochem. Int."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-94-017-8914-1_1","article-title":"How can we identify ictal and interictal abnormal activity?","volume":"813","author":"Fisher","year":"2014","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S0013-4694(98)00051-0","article-title":"Electrophysiological correlates of sleep delta waves","volume":"107","author":"Amzica","year":"1998","journal-title":"Electroencephalogr. Clin. Neurophysiol."},{"key":"ref_45","unstructured":"Bear, M.F., Connors, B.W., and Paradiso, M.A. (2007). Neuroscience, Lippincott Williams & Wilkins. [3rd ed.]."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1111\/j.1528-1157.1997.tb01097.x","article-title":"Evolution of epilepsy and EEG findings in Angelman syndrome","volume":"38","author":"Laan","year":"1997","journal-title":"Epilepsia"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/0013-4694(90)90134-6","article-title":"Asymmetry in delta activity in patients with focal epilepsy","volume":"75","author":"Gotman","year":"1990","journal-title":"Electroencephalogr. Clin. Neurophysiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1007\/s10072-003-0183-2","article-title":"Epileptic seizures in multiple sclerosis: Clinical and EEG correlations","volume":"24","author":"Striano","year":"2003","journal-title":"Neurol. Sci."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/20\/5903\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:24:08Z","timestamp":1760178248000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/20\/5903"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,19]]},"references-count":48,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["s20205903"],"URL":"https:\/\/doi.org\/10.3390\/s20205903","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,19]]}}}