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Sperm morphological analysis plays an important role in assessing sperm fertility. However, current sperm morphological analysis is usually performed using microscopic images of stained sperm, which requires complex staining operations, making it difficult to achieve a comprehensive and efficient diagnosis. In this work, we propose and experimentally demonstrate sperm morphological analysis with optofluidic time\u2010stretch (OTS) imaging flow cytometry. Specifically, we capture the stain\u2010free sperm images with high\u2010throughput OTS imaging flow cytometry and analyze the morphological features using a U\u2010Net network. The analysis of 40,000 images from 20 clinical semen samples indicates that our method achieves higher accuracy than the smear test and shows high concordance with the results using comprehensive methods of routine semen examination. This work provides a high\u2010throughput, stain\u2010free sperm morphological analysis method, which holds promise for a comprehensive evaluation of semen.<\/jats:p>","DOI":"10.1002\/jbio.202400560","type":"journal-article","created":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T05:31:09Z","timestamp":1741757469000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["High\u2010Throughput and Stain\u2010Free Morphological Analysis of Sperm Using Optofluidic Time\u2010Stretch Imaging Flow Cytometry"],"prefix":"10.1002","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8910-8904","authenticated-orcid":false,"given":"Yueyun","family":"Weng","sequence":"first","affiliation":[{"name":"Institute of Technological Sciences, Wuhan University  Wuhan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Zou","sequence":"additional","affiliation":[{"name":"Reproductive Medical 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