Precision isolation and cultivation of single cells by vortex and flat-top laser ejection
Single-cell isolation stands as a critical step in single-cell studies, and single-cell ejection technology based on laser induced forward transfer technology (LIFT) is considered one of the most promising methods in this regard for its ability of visible isolating single cell from complex samples....
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2024-04-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1369506/full |
| _version_ | 1797215838614323200 |
|---|---|
| author | Fuyuan Chen Fuyuan Chen Kunxiang Liu Kunxiang Liu Lindong Shang Lindong Shang Yuntong Wang Yuntong Wang Xusheng Tang Xusheng Tang Peng Liang Peng Liang Bei Li Bei Li |
| author_facet | Fuyuan Chen Fuyuan Chen Kunxiang Liu Kunxiang Liu Lindong Shang Lindong Shang Yuntong Wang Yuntong Wang Xusheng Tang Xusheng Tang Peng Liang Peng Liang Bei Li Bei Li |
| author_sort | Fuyuan Chen |
| collection | DOAJ |
| description | Single-cell isolation stands as a critical step in single-cell studies, and single-cell ejection technology based on laser induced forward transfer technology (LIFT) is considered one of the most promising methods in this regard for its ability of visible isolating single cell from complex samples. In this study, we improve the LIFT technology and introduce optical vortex laser-induced forward transfer (OV-LIFT) and flat-top laser-induced forward transfer (FT-LIFT) by utilizing spatial light modulator (SLM), aiming to enhance the precision of single-cell sorting and the cell’s viability after ejection. Experimental results demonstrate that applying vortex and flat-top beams during the sorting and collection process enables precise retrieval of single cells within diameter ranges of 50 μm and 100 μm, respectively. The recovery rates of Saccharomyces cerevisiae and Escherichia coli DH5α single cell ejected by vortex beam are 89 and 78%, by flat-top beam are 85 and 57%. When employing Gaussian beam sorting, the receiving range extends to 400 μm, with cultivation success rates of S. cerevisiae and E. coli DH5α single cell are 48 and 19%, respectively. This marks the first application of different mode beams in the ejection and cultivation of single cells, providing a novel and effective approach for the precise isolation and improving the viability of single cells. |
| first_indexed | 2024-04-24T11:36:26Z |
| format | Article |
| id | doaj.art-9953a9ddfd1a499f932f86f979b7ac0a |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-24T11:36:26Z |
| publishDate | 2024-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-9953a9ddfd1a499f932f86f979b7ac0a2024-04-10T05:16:07ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-04-011510.3389/fmicb.2024.13695061369506Precision isolation and cultivation of single cells by vortex and flat-top laser ejectionFuyuan Chen0Fuyuan Chen1Kunxiang Liu2Kunxiang Liu3Lindong Shang4Lindong Shang5Yuntong Wang6Yuntong Wang7Xusheng Tang8Xusheng Tang9Peng Liang10Peng Liang11Bei Li12Bei Li13Key Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaSingle-cell isolation stands as a critical step in single-cell studies, and single-cell ejection technology based on laser induced forward transfer technology (LIFT) is considered one of the most promising methods in this regard for its ability of visible isolating single cell from complex samples. In this study, we improve the LIFT technology and introduce optical vortex laser-induced forward transfer (OV-LIFT) and flat-top laser-induced forward transfer (FT-LIFT) by utilizing spatial light modulator (SLM), aiming to enhance the precision of single-cell sorting and the cell’s viability after ejection. Experimental results demonstrate that applying vortex and flat-top beams during the sorting and collection process enables precise retrieval of single cells within diameter ranges of 50 μm and 100 μm, respectively. The recovery rates of Saccharomyces cerevisiae and Escherichia coli DH5α single cell ejected by vortex beam are 89 and 78%, by flat-top beam are 85 and 57%. When employing Gaussian beam sorting, the receiving range extends to 400 μm, with cultivation success rates of S. cerevisiae and E. coli DH5α single cell are 48 and 19%, respectively. This marks the first application of different mode beams in the ejection and cultivation of single cells, providing a novel and effective approach for the precise isolation and improving the viability of single cells.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1369506/fulllaser induced forward transfervortex beamflat-top beamsingle cell sortingsingle-cell culture |
| spellingShingle | Fuyuan Chen Fuyuan Chen Kunxiang Liu Kunxiang Liu Lindong Shang Lindong Shang Yuntong Wang Yuntong Wang Xusheng Tang Xusheng Tang Peng Liang Peng Liang Bei Li Bei Li Precision isolation and cultivation of single cells by vortex and flat-top laser ejection Frontiers in Microbiology laser induced forward transfer vortex beam flat-top beam single cell sorting single-cell culture |
| title | Precision isolation and cultivation of single cells by vortex and flat-top laser ejection |
| title_full | Precision isolation and cultivation of single cells by vortex and flat-top laser ejection |
| title_fullStr | Precision isolation and cultivation of single cells by vortex and flat-top laser ejection |
| title_full_unstemmed | Precision isolation and cultivation of single cells by vortex and flat-top laser ejection |
| title_short | Precision isolation and cultivation of single cells by vortex and flat-top laser ejection |
| title_sort | precision isolation and cultivation of single cells by vortex and flat top laser ejection |
| topic | laser induced forward transfer vortex beam flat-top beam single cell sorting single-cell culture |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1369506/full |
| work_keys_str_mv | AT fuyuanchen precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT fuyuanchen precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT kunxiangliu precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT kunxiangliu precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT lindongshang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT lindongshang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT yuntongwang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT yuntongwang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT xushengtang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT xushengtang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT pengliang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT pengliang precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT beili precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection AT beili precisionisolationandcultivationofsinglecellsbyvortexandflattoplaserejection |