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The viability of isolated cells is the key prerequisite for the downstream molecular analysis, especially for RNA sequencing. This study develops a laser\u2010induced forward transfer \u2010assisted microfiltration system (LIFT\u2010AMFS) for high\u2010viability CTC enrichment and retrieval from whole blood. The LIFT\u2010compatible double\u2010stepped microfilter (DSMF), central to this system, comprises two micropore layers: the lower layer's smaller micropores facilitate size\u2010based cell separation, and the upper layer's larger micropores enable liquid encapsulating captured cells. By optimizing the design of the DSMFs, the system has a capture efficiency of 88% at the processing throughput of up to 15.0\u00a0mL\u00a0min\n                    <jats:sup>\u22121<\/jats:sup>\n                    during the microfilter\u2010based size screening stage, with a single\u2010cell yield of over 95% during the retrieval stage. The retrieved single cells, with high viability, are qualified for ex vivo culture and direct RNA sequencing. The cDNA yield from isolated CTCs surpassed 4.5\u00a0ng, sufficient for library construction. All single\u2010cell sequencing data exhibited Q30 scores above 95.92%. The LIFT\u2010AMFS shows promise in cellular and biomedical research.\n                  <\/jats:p>","DOI":"10.1002\/advs.202414195","type":"journal-article","created":{"date-parts":[[2025,1,27]],"date-time":"2025-01-27T08:46:04Z","timestamp":1737967564000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["High\u2010Viability Circulating Tumor Cells Sorting From Whole Blood at Single Cell Level Using Laser\u2010Induced Forward Transfer\u2010Assisted Microfiltration"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6685-5082","authenticated-orcid":false,"given":"Qingmei","family":"Xu","sequence":"first","affiliation":[{"name":"School of Integrated Circuits Peking University  Beijing 100871 China"},{"name":"Department of Electrical Engineering Taiyuan Institute of Technology  Taiyuan 030008 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4911-9446","authenticated-orcid":false,"given":"Yuntong","family":"Wang","sequence":"additional","affiliation":[{"name":"Changchun Institute of Optics Fine Mechanics and Physics Chinese Academy of Sciences  Changchun 130033 China"},{"name":"University of Chinese Academy of Sciences  Beijing 100049 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Songtao","family":"Dou","sequence":"additional","affiliation":[{"name":"School of Integrated Circuits Peking University  Beijing 100871 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Xu","sequence":"additional","affiliation":[{"name":"Cancer Translational Medicine Research Center The Second Affiliated Hospital of Dalian Medical University  Dalian 116027 China"},{"name":"Department of Respiratory Medicine The Second Affiliated Hospital of Dalian Medical University  Dalian 116027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2639-6181","authenticated-orcid":false,"given":"Zhenhe","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism East China University of Science and Technology  Shanghai 200237 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2742-5354","authenticated-orcid":false,"given":"Han","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Integrated Circuits Peking University  Beijing 100871 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Integrated Circuits Peking University  Beijing 100871 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanming","family":"Xia","sequence":"additional","affiliation":[{"name":"Guangzhou National Laboratory  Guangzhou 510320 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Xue","sequence":"additional","affiliation":[{"name":"Hooke Laboratory  Changchun 130033 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hang","family":"Li","sequence":"additional","affiliation":[{"name":"Hooke Laboratory  Changchun 130033 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7260-9504","authenticated-orcid":false,"given":"Xiao","family":"Ma","sequence":"additional","affiliation":[{"name":"Hangzhou Branemagic Medical Technology Co. 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