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Imaging flow cytometry, in particular, enables large-scale image-based profiling of heterogeneous cell populations, allowing statistical analysis of cellular morphology, subcellular features, and functional responses. However, its analytical capability is often limited by the use of conventional two-dimensional (2D) image sensors. In this review, we highlight recent advances in single-pixel imaging flow cytometry, which replaces 2D image sensors with single-pixel photodetectors. This approach offers advantages in sensitivity, flexibility, and speed in imaging system design and has been implemented in various optical configurations to achieve high-throughput single-cell imaging. We first introduce its key techniques, then outline representative biomedical applications, including cancer and COVID-19 research, and finally discuss current limitations and prospects for future developments. Single-pixel imaging flow cytometry is expected to serve as a versatile platform supporting both basic and translational studies in diverse biomedical applications.<\/jats:p>","DOI":"10.1186\/s41232-025-00401-5","type":"journal-article","created":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T05:26:18Z","timestamp":1765517178000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Single-pixel imaging flow cytometry for biomedical research"],"prefix":"10.1186","volume":"45","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3340-9462","authenticated-orcid":false,"given":"Hiroshi","family":"Kanno","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiying","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rio","family":"Sato","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidenori","family":"Endo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuniyasu","family":"Niizuma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6302-6038","authenticated-orcid":false,"given":"Keisuke","family":"Goda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,12]]},"reference":[{"key":"401_CR1","doi-asserted-by":"publisher","first-page":"618","DOI":"10.1038\/nrg3542","volume":"14","author":"E Shapiro","year":"2013","unstructured":"Shapiro E, Biezuner T, Linnarsson S. 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