Multi-step processing of single cells using semi-permeable capsules
| العنوان: | Multi-step processing of single cells using semi-permeable capsules |
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| المؤلفون: | Karolis Leonavicius, Rolandas Meškys, Greta Leonaviciene, Linas Mazutis |
| المصدر: | Lab on a Chip. 20:4052-4062 |
| بيانات النشر: | Royal Society of Chemistry (RSC), 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Phenotypic screening, Microfluidics, Biomedical Engineering, Capsules, Bioengineering, Nanotechnology, 02 engineering and technology, Genome amplification, Biochemistry, Bacterial cell structure, 03 medical and health sciences, Semipermeable membrane, Droplet microfluidics, Molecular Biology, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Bacteria, Biomolecule, Hydrogels, General Chemistry, 021001 nanoscience & nanotechnology, chemistry, Self-healing hydrogels, 0210 nano-technology |
| الوصف: | Droplet microfluidics technology provides a powerful approach to isolate and process millions of single cells simultaneously. Despite many exciting applications that have emerged based on this technology, workflows based on multi-step operations, including molecular biology and cell-based phenotypic screening assays, cannot be easily adapted to droplet format. Here, we present a microfluidics-based technique to isolate single cells, or biological samples, into semi-permeable hydrogel capsules and perform multi-step biological workflows on thousands to millions of individual cells simultaneously. The biochemical reactions are performed by changing the aqueous buffer surrounding the capsules, without needing sophisticated equipment. The semi-permeable nature of the capsules' shell retains large encapsulated biomolecules (such as genome) while allowing smaller molecules (such as proteins) to passively diffuse. In contrast to conventional hydrogel bead assays, the approach presented here improves bacterial cell retention during multi-step procedures as well as the efficiency of biochemical reactions. We showcase two examples of capsule use for single genome amplification of bacteria, and expansion of individual clones into isogenic microcolonies for later screening for biodegradable plastic production. |
| تدمد: | 1473-0189 1473-0197 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::43d19bc8e1f746d61ec16fd0840b04c4 https://doi.org/10.1039/d0lc00660b |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....43d19bc8e1f746d61ec16fd0840b04c4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14730189 14730197 |
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