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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,...

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Autores principales: Tewari Kumar, Phalguni, Decrop, Deborah, Safdar, Saba, Passaris, Ioannis, Kokalj, Tadej, Puers, Robert, Aertsen, Abram, Spasic, Dragana, Lammertyn, Jeroen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7142809/
https://www.ncbi.nlm.nih.gov/pubmed/32183431
http://dx.doi.org/10.3390/mi11030308
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author Tewari Kumar, Phalguni
Decrop, Deborah
Safdar, Saba
Passaris, Ioannis
Kokalj, Tadej
Puers, Robert
Aertsen, Abram
Spasic, Dragana
Lammertyn, Jeroen
author_facet Tewari Kumar, Phalguni
Decrop, Deborah
Safdar, Saba
Passaris, Ioannis
Kokalj, Tadej
Puers, Robert
Aertsen, Abram
Spasic, Dragana
Lammertyn, Jeroen
author_sort Tewari Kumar, Phalguni
collection PubMed
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 Salmonella Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent S. 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.
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spelling pubmed-71428092020-04-14 Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers Tewari Kumar, Phalguni Decrop, Deborah Safdar, Saba Passaris, Ioannis Kokalj, Tadej Puers, Robert Aertsen, Abram Spasic, Dragana Lammertyn, Jeroen Micromachines (Basel) Article 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 Salmonella Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent S. 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. MDPI 2020-03-15 /pmc/articles/PMC7142809/ /pubmed/32183431 http://dx.doi.org/10.3390/mi11030308 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tewari Kumar, Phalguni
Decrop, Deborah
Safdar, Saba
Passaris, Ioannis
Kokalj, Tadej
Puers, Robert
Aertsen, Abram
Spasic, Dragana
Lammertyn, Jeroen
Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7142809/
https://www.ncbi.nlm.nih.gov/pubmed/32183431
http://dx.doi.org/10.3390/mi11030308
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