Selective single-bacteria extraction based on capture and release of microemulsion droplets
Human host-associated microbial communities in body sites can reflect health status based on the population distribution and specific microbial properties in the heterogeneous community. Bacteria identification at the single-cell level provides a reliable biomarker and pathological information for c...
| Main Authors: | , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2023
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/171227 |
| _version_ | 1811696199063830528 |
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| author | Li, Jiyu Hu, Dinglong Lim, Chee Kent Ren, Jifeng Yao, Xin Ma, Chao Marcos Chen, Weiqiang Lee, Patrick K. H. Lam, Raymond H. W. |
| author2 | School of Mechanical and Aerospace Engineering |
| author_facet | School of Mechanical and Aerospace Engineering Li, Jiyu Hu, Dinglong Lim, Chee Kent Ren, Jifeng Yao, Xin Ma, Chao Marcos Chen, Weiqiang Lee, Patrick K. H. Lam, Raymond H. W. |
| author_sort | Li, Jiyu |
| collection | NTU |
| description | Human host-associated microbial communities in body sites can reflect health status based on the population distribution and specific microbial properties in the heterogeneous community. Bacteria identification at the single-cell level provides a reliable biomarker and pathological information for clinical diagnosis. Nevertheless, biosamples obtained from some body sites cannot offer sufficient sample volume and number of target cells as required by most of the existing single-cell isolation methods such as flow cytometry. Herein we report a novel integrated microfluidic system, which consists of a microemulsion module for single-bacteria encapsulation and a sequential microdroplet capture and release module for selectively extracting only the single-bacteria encapsulated in microdroplets. We optimize the system for a success rate of the single-cell extraction to be > 38%. We further verify applicability of the system with prepared cell mixtures (Methylorubrum extorquens AM1 and Methylomicrobium album BG8) and biosamples collected from human skin, to quantify the population distribution of multiple key species in a heterogeneous microbial community. Results indicate perfect viability of the single-cell extracts and compatibility with downstream analyses such as PCR. Together, this research demonstrates that the reported single-bacteria extraction system can be applied in microbiome and pathology research and clinical diagnosis as a clinical or point-of-care device. |
| first_indexed | 2024-10-01T07:35:34Z |
| format | Journal Article |
| id | ntu-10356/171227 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:35:34Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | ntu-10356/1712272023-10-21T16:48:05Z Selective single-bacteria extraction based on capture and release of microemulsion droplets Li, Jiyu Hu, Dinglong Lim, Chee Kent Ren, Jifeng Yao, Xin Ma, Chao Marcos Chen, Weiqiang Lee, Patrick K. H. Lam, Raymond H. W. School of Mechanical and Aerospace Engineering Science::Biological sciences Cell Separation Bacteria Human host-associated microbial communities in body sites can reflect health status based on the population distribution and specific microbial properties in the heterogeneous community. Bacteria identification at the single-cell level provides a reliable biomarker and pathological information for clinical diagnosis. Nevertheless, biosamples obtained from some body sites cannot offer sufficient sample volume and number of target cells as required by most of the existing single-cell isolation methods such as flow cytometry. Herein we report a novel integrated microfluidic system, which consists of a microemulsion module for single-bacteria encapsulation and a sequential microdroplet capture and release module for selectively extracting only the single-bacteria encapsulated in microdroplets. We optimize the system for a success rate of the single-cell extraction to be > 38%. We further verify applicability of the system with prepared cell mixtures (Methylorubrum extorquens AM1 and Methylomicrobium album BG8) and biosamples collected from human skin, to quantify the population distribution of multiple key species in a heterogeneous microbial community. Results indicate perfect viability of the single-cell extracts and compatibility with downstream analyses such as PCR. Together, this research demonstrates that the reported single-bacteria extraction system can be applied in microbiome and pathology research and clinical diagnosis as a clinical or point-of-care device. Published version We acknowledge financial supports from Hong Kong Research Grant Council (General Research Grant 11216220), National Natural Science Foundation of China (NSFC Grant 31770920) and City University of Hong Kong (Strategic Interdisciplinary Research Grant 7020049). 2023-10-20T02:23:43Z 2023-10-20T02:23:43Z 2022 Journal Article Li, J., Hu, D., Lim, C. K., Ren, J., Yao, X., Ma, C., Marcos, Chen, W., Lee, P. K. H. & Lam, R. H. W. (2022). Selective single-bacteria extraction based on capture and release of microemulsion droplets. Scientific Reports, 12(1), 15461-. https://dx.doi.org/10.1038/s41598-022-19844-8 2045-2322 https://hdl.handle.net/10356/171227 10.1038/s41598-022-19844-8 36104374 2-s2.0-85137921070 1 12 15461 en Scientific Reports © 2022 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. application/pdf |
| spellingShingle | Science::Biological sciences Cell Separation Bacteria Li, Jiyu Hu, Dinglong Lim, Chee Kent Ren, Jifeng Yao, Xin Ma, Chao Marcos Chen, Weiqiang Lee, Patrick K. H. Lam, Raymond H. W. Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title | Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title_full | Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title_fullStr | Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title_full_unstemmed | Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title_short | Selective single-bacteria extraction based on capture and release of microemulsion droplets |
| title_sort | selective single bacteria extraction based on capture and release of microemulsion droplets |
| topic | Science::Biological sciences Cell Separation Bacteria |
| url | https://hdl.handle.net/10356/171227 |
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