A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay
A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single inf...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2013
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| Online Access: | https://hdl.handle.net/10356/102469 http://hdl.handle.net/10220/16867 |
| _version_ | 1826109965191872512 |
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| author | Zhang, Rui Gong Haiqing, Thomas Zeng, Xu Dong Sze, Chun Chau |
| author2 | School of Mechanical and Aerospace Engineering |
| author_facet | School of Mechanical and Aerospace Engineering Zhang, Rui Gong Haiqing, Thomas Zeng, Xu Dong Sze, Chun Chau |
| author_sort | Zhang, Rui |
| collection | NTU |
| description | A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single infected host cells into the individual micro-wells of the biochip was achieved by one-step vacuum-driven microfluidics. The adhered bacterial cells were then quantified by direct on-chip real-time PCR assay with single-bacterium-detection sensitivity. The performance of this microfluidic platform was demonstrated through profiling of the association of a common bacterial pathogen, Pseudomonas aeruginosa, to single host human lung epithelial A549 cells, revealing an adherence distribution that has not been previously reported. This microfluidic platform offers a simple and effective tool for biologists to analyze pathogen–host interaction at the single-cell level without the necessities of fluorescence labeling. The chip can similarly be used for other PCR-based applications requiring single-cell analysis. |
| first_indexed | 2024-10-01T02:26:43Z |
| format | Journal Article |
| id | ntu-10356/102469 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:26:43Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/1024692020-03-07T13:22:20Z A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay Zhang, Rui Gong Haiqing, Thomas Zeng, Xu Dong Sze, Chun Chau School of Mechanical and Aerospace Engineering School of Biological Sciences Nanyang Environment and Water Research Institute DRNTU::Science::Biological sciences A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single infected host cells into the individual micro-wells of the biochip was achieved by one-step vacuum-driven microfluidics. The adhered bacterial cells were then quantified by direct on-chip real-time PCR assay with single-bacterium-detection sensitivity. The performance of this microfluidic platform was demonstrated through profiling of the association of a common bacterial pathogen, Pseudomonas aeruginosa, to single host human lung epithelial A549 cells, revealing an adherence distribution that has not been previously reported. This microfluidic platform offers a simple and effective tool for biologists to analyze pathogen–host interaction at the single-cell level without the necessities of fluorescence labeling. The chip can similarly be used for other PCR-based applications requiring single-cell analysis. 2013-10-25T01:36:55Z 2019-12-06T20:55:28Z 2013-10-25T01:36:55Z 2019-12-06T20:55:28Z 2013 2013 Journal Article Zhang, R., Gong, H. T., Zeng, X. D., & Sze, C. C. (2013). A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay. Analytical and bioanalytical chemistry, 405(12), 4277-4282. 1618-2642 https://hdl.handle.net/10356/102469 http://hdl.handle.net/10220/16867 10.1007/s00216-013-6826-0 en Analytical and bioanalytical chemistry |
| spellingShingle | DRNTU::Science::Biological sciences Zhang, Rui Gong Haiqing, Thomas Zeng, Xu Dong Sze, Chun Chau A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title | A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title_full | A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title_fullStr | A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title_full_unstemmed | A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title_short | A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay |
| title_sort | high throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real time pcr assay |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/102469 http://hdl.handle.net/10220/16867 |
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