DC Field | Value | Language |
---|---|---|
dc.contributor.author | Choi, Jong-Seob | ko |
dc.contributor.author | Bae, Sunwoong | ko |
dc.contributor.author | Kim, Kyung Hoon | ko |
dc.contributor.author | Kim, Jaoon Y. H. | ko |
dc.contributor.author | Sim, Sang Jun | ko |
dc.contributor.author | Seo, Tae-Seok | ko |
dc.date.accessioned | 2015-11-20T09:12:54Z | - |
dc.date.available | 2015-11-20T09:12:54Z | - |
dc.date.created | 2015-01-05 | - |
dc.date.created | 2015-01-05 | - |
dc.date.issued | 2014 | - |
dc.identifier.citation | RSC ADVANCES, v.4, no.106, pp.61298 - 61304 | - |
dc.identifier.issn | 2046-2069 | - |
dc.identifier.uri | http://hdl.handle.net/10203/201090 | - |
dc.description.abstract | A perforated hemispherical microwell structure is shown to efficiently capture single Chlamydomonas reinhardtii (C. reinhardtii) cells, culture them to form colonies, and retrieve these colonies to serve as seeds for large-scale cultivation. This solution-phase formation and recovery of colonies could overcome the tedious and time-consuming process of selecting colonies from a solid-phase agar plate. The fabricated microdevice was composed of three layers: a top layer consisting of a cell solution for injection and recovery of a microalgal solution, a hemispherical perforated microwell array in the middle, and a bottom layer in which the solution is manipulated by controlling the hydrodynamic force. The microalgal (wild type and hygromycin B-resistant mutant) cells loaded in the top layer rapidly diffused into the microwell holes, and individual such cells were captured with high efficiency (>90%) and within 1 min by applying a withdraw mode in the bottom layer. Single-cell-based cultivation in a medium containing hygromycin B was then performed to generate colonies in the hemispherical microwell. While the wild type cells died, mutant cells resistant to hygromycin B survived well and grew into a colony within 2 days. The produced colonies in the microwells were recovered by applying a release mode in the bottom layer, so that a hydrodynamic force was exerted vertically to push out the colonies through the outlet in 10 s. The recovered cells were cultured on a large scale in medium by using a flask. The recovered C. reinhardtii was confirmed as a hygromycin B-resistant mutant by identifying the hygromycin gene in the polymerase chain reaction (PCR). The microdevice described here could in solution perform single-cell capture, colony formation, and retrieval of colonies for further large-scale cultivation, which could replace tedious and time-consuming solid-phase agar plate processes with a 7-fold reduction in the duration of the process. | - |
dc.language | English | - |
dc.publisher | ROYAL SOC CHEMISTRY | - |
dc.subject | CHLAMYDOMONAS-REINHARDTII | - |
dc.subject | ARRAYS | - |
dc.title | Capture and culturing of single microalgae cells, and retrieval of colonies using a perforated hemispherical microwell structure | - |
dc.type | Article | - |
dc.identifier.wosid | 000345656600008 | - |
dc.identifier.scopusid | 2-s2.0-84913559533 | - |
dc.type.rims | ART | - |
dc.citation.volume | 4 | - |
dc.citation.issue | 106 | - |
dc.citation.beginningpage | 61298 | - |
dc.citation.endingpage | 61304 | - |
dc.citation.publicationname | RSC ADVANCES | - |
dc.identifier.doi | 10.1039/c4ra09730k | - |
dc.contributor.localauthor | Seo, Tae-Seok | - |
dc.contributor.nonIdAuthor | Kim, Jaoon Y. H. | - |
dc.contributor.nonIdAuthor | Sim, Sang Jun | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordPlus | CHLAMYDOMONAS-REINHARDTII | - |
dc.subject.keywordPlus | ARRAYS | - |
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