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They swiftly respond to vascular injury by adhering to damaged vessel surfaces, activating signalling pathways, and aggregating with each other to control bleeding. This dynamic process of platelet activation is intricately coordinated, spanning from membrane receptor maturation to intracellular interactions to whole-cell responses. Live-cell imaging has become an invaluable tool for dissecting these complexes. Despite its benefits, live imaging of platelets presents significant technical challenges. This review addresses these challenges, identifying key areas in need of further development and proposing possible solutions. We also focus on the dynamic processes of platelet adhesion, activation, and aggregation in haemostasis and thrombosis, applying imaging capacities from the microscale to the nanoscale. By exploring various live imaging techniques, we demonstrate how these approaches offer crucial insights into platelet biology and deepen our understanding of these three core events. In conclusion, this review provides an overview of the imaging methods currently available for studying platelet dynamics, guiding researchers in selecting suitable techniques for specific studies. By advancing our knowledge of platelet behaviour, these imaging methods contribute to research on haemostasis, thrombosis, and platelet-related diseases, ultimately aiming to improve clinical outcomes.<\/jats:p>","DOI":"10.3390\/s25020491","type":"journal-article","created":{"date-parts":[[2025,1,16]],"date-time":"2025-01-16T06:46:10Z","timestamp":1737009970000},"page":"491","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Advancing Platelet Research Through Live-Cell Imaging: Challenges, Techniques, and Insights"],"prefix":"10.3390","volume":"25","author":[{"given":"Yuping Yolanda","family":"Tan","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia"},{"name":"Heart Research Institute, Newtown, NSW 2042, Australia"},{"name":"Charles Perkins Centre, The University of Sydney, Camperdown, NSW 2006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinghan","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia"},{"name":"Heart Research Institute, Newtown, NSW 2042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7364-3945","authenticated-orcid":false,"given":"Qian Peter","family":"Su","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia"},{"name":"Heart Research Institute, Newtown, NSW 2042, Australia"},{"name":"Charles Perkins Centre, The University of Sydney, Camperdown, NSW 2006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,16]]},"reference":[{"key":"ref_1","first-page":"e55658","article-title":"Live-cell imaging of platelet degranulation and secretion under flow","volume":"125","author":"Barendrecht","year":"2017","journal-title":"J. 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