Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers
When screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former,...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-03-01
|
| Series: | Micromachines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-666X/11/3/308 |
| _version_ | 1828968502003761152 |
|---|---|
| author | Phalguni Tewari Kumar Deborah Decrop Saba Safdar Ioannis Passaris Tadej Kokalj Robert Puers Abram Aertsen Dragana Spasic Jeroen Lammertyn |
| author_facet | Phalguni Tewari Kumar Deborah Decrop Saba Safdar Ioannis Passaris Tadej Kokalj Robert Puers Abram Aertsen Dragana Spasic Jeroen Lammertyn |
| author_sort | Phalguni Tewari Kumar |
| collection | DOAJ |
| description | When screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former, although allowing cell sorting, fails to track dynamic cell behavior, while the latter has been limited to complex channel-based microfluidic platforms. In this study, digital microfluidics (DMF) was integrated with OT for selective trapping, relocation, and further proliferation of single bacterial cells, while offering continuous imaging of cells to evaluate dynamic cell behavior. To enable this, magnetic beads coated with <i>Salmonella</i> Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent <i>S.</i> Typhimurium population. Next, OT were used to select a bead with a bacterium of interest, based on its fluorescent expression, and to relocate this bead to a different microwell on the same or different array. Using an agar patch affixed on top, the relocated bacterium was subsequently allowed to proliferate. Our OT-integrated DMF platform thus successfully enabled selective trapping, retrieval, relocation, and proliferation of bacteria of interest at single-cell level, thereby enabling their downstream analysis. |
| first_indexed | 2024-12-14T12:12:23Z |
| format | Article |
| id | doaj.art-7261beadb7be42c68e8c3f4f99495354 |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-12-14T12:12:23Z |
| publishDate | 2020-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj.art-7261beadb7be42c68e8c3f4f994953542022-12-21T23:01:42ZengMDPI AGMicromachines2072-666X2020-03-0111330810.3390/mi11030308mi11030308Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical TweezersPhalguni Tewari Kumar0Deborah Decrop1Saba Safdar2Ioannis Passaris3Tadej Kokalj4Robert Puers5Abram Aertsen6Dragana Spasic7Jeroen Lammertyn8Department of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumDepartment of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumDepartment of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumDepartment of Microbial and Molecular Systems, KU Leuven, 3001 Leuven, BelgiumDepartment of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumESAT-MICAS, Microelectronics and Sensors, KU Leuven, 3001 Leuven, BelgiumDepartment of Microbial and Molecular Systems, KU Leuven, 3001 Leuven, BelgiumDepartment of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumDepartment of Biosystems, Biosensors Group, KU Leuven, 3001 Leuven, BelgiumWhen screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former, although allowing cell sorting, fails to track dynamic cell behavior, while the latter has been limited to complex channel-based microfluidic platforms. In this study, digital microfluidics (DMF) was integrated with OT for selective trapping, relocation, and further proliferation of single bacterial cells, while offering continuous imaging of cells to evaluate dynamic cell behavior. To enable this, magnetic beads coated with <i>Salmonella</i> Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent <i>S.</i> Typhimurium population. Next, OT were used to select a bead with a bacterium of interest, based on its fluorescent expression, and to relocate this bead to a different microwell on the same or different array. Using an agar patch affixed on top, the relocated bacterium was subsequently allowed to proliferate. Our OT-integrated DMF platform thus successfully enabled selective trapping, retrieval, relocation, and proliferation of bacteria of interest at single-cell level, thereby enabling their downstream analysis.https://www.mdpi.com/2072-666X/11/3/308optical tweezerssingle-celldigital microfluidics<i>salmonella</i> typhimurium |
| spellingShingle | Phalguni Tewari Kumar Deborah Decrop Saba Safdar Ioannis Passaris Tadej Kokalj Robert Puers Abram Aertsen Dragana Spasic Jeroen Lammertyn Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers Micromachines optical tweezers single-cell digital microfluidics <i>salmonella</i> typhimurium |
| title | Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers |
| title_full | Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers |
| title_fullStr | Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers |
| title_full_unstemmed | Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers |
| title_short | Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers |
| title_sort | digital microfluidics for single bacteria capture and selective retrieval using optical tweezers |
| topic | optical tweezers single-cell digital microfluidics <i>salmonella</i> typhimurium |
| url | https://www.mdpi.com/2072-666X/11/3/308 |
| work_keys_str_mv | AT phalgunitewarikumar digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT deborahdecrop digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT sabasafdar digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT ioannispassaris digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT tadejkokalj digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT robertpuers digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT abramaertsen digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT draganaspasic digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers AT jeroenlammertyn digitalmicrofluidicsforsinglebacteriacaptureandselectiveretrievalusingopticaltweezers |