Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli
Lipopolysaccharide (LPS) is the unique feature that composes the outer leaflet of the Gram-negative bacterial cell envelope. Variations in LPS structures affect a number of physiological processes, including outer membrane permeability, antimicrobial resistance, recognition by the host immune system...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-02-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.991784/full |
| _version_ | 1797904688335552512 |
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| author | Qianru Wang Hyungseok Kim Tiffany M. Halvorsen Sijie Chen Christopher S. Hayes Cullen R. Buie |
| author_facet | Qianru Wang Hyungseok Kim Tiffany M. Halvorsen Sijie Chen Christopher S. Hayes Cullen R. Buie |
| author_sort | Qianru Wang |
| collection | DOAJ |
| description | Lipopolysaccharide (LPS) is the unique feature that composes the outer leaflet of the Gram-negative bacterial cell envelope. Variations in LPS structures affect a number of physiological processes, including outer membrane permeability, antimicrobial resistance, recognition by the host immune system, biofilm formation, and interbacterial competition. Rapid characterization of LPS properties is crucial for studying the relationship between these LPS structural changes and bacterial physiology. However, current assessments of LPS structures require LPS extraction and purification followed by cumbersome proteomic analysis. This paper demonstrates one of the first high-throughput and non-invasive strategies to directly distinguish Escherichia coli with different LPS structures. Using a combination of three-dimensional insulator-based dielectrophoresis (3DiDEP) and cell tracking in a linear electrokinetics assay, we elucidate the effect of structural changes in E. coli LPS oligosaccharides on electrokinetic mobility and polarizability. We show that our platform is sufficiently sensitive to detect LPS structural variations at the molecular level. To correlate electrokinetic properties of LPS with the outer membrane permeability, we further examined effects of LPS structural variations on bacterial susceptibility to colistin, an antibiotic known to disrupt the outer membrane by targeting LPS. Our results suggest that microfluidic electrokinetic platforms employing 3DiDEP can be a useful tool for isolating and selecting bacteria based on their LPS glycoforms. Future iterations of these platforms could be leveraged for rapid profiling of pathogens based on their surface LPS structural identity. |
| first_indexed | 2024-04-10T09:52:57Z |
| format | Article |
| id | doaj.art-336042ec0fe34beabbc29903152b160d |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-10T09:52:57Z |
| publishDate | 2023-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-336042ec0fe34beabbc29903152b160d2023-02-16T13:49:37ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-02-011110.3389/fbioe.2023.991784991784Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coliQianru Wang0Hyungseok Kim1Tiffany M. Halvorsen2Sijie Chen3Christopher S. Hayes4Cullen R. Buie5Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United StatesDepartment of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United StatesDepartment of Molecular, Cellular and Developmental Biology, University of California, Santa Barbara, Santa Barbara, CA, United StatesDepartment of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United StatesDepartment of Molecular, Cellular and Developmental Biology, University of California, Santa Barbara, Santa Barbara, CA, United StatesDepartment of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United StatesLipopolysaccharide (LPS) is the unique feature that composes the outer leaflet of the Gram-negative bacterial cell envelope. Variations in LPS structures affect a number of physiological processes, including outer membrane permeability, antimicrobial resistance, recognition by the host immune system, biofilm formation, and interbacterial competition. Rapid characterization of LPS properties is crucial for studying the relationship between these LPS structural changes and bacterial physiology. However, current assessments of LPS structures require LPS extraction and purification followed by cumbersome proteomic analysis. This paper demonstrates one of the first high-throughput and non-invasive strategies to directly distinguish Escherichia coli with different LPS structures. Using a combination of three-dimensional insulator-based dielectrophoresis (3DiDEP) and cell tracking in a linear electrokinetics assay, we elucidate the effect of structural changes in E. coli LPS oligosaccharides on electrokinetic mobility and polarizability. We show that our platform is sufficiently sensitive to detect LPS structural variations at the molecular level. To correlate electrokinetic properties of LPS with the outer membrane permeability, we further examined effects of LPS structural variations on bacterial susceptibility to colistin, an antibiotic known to disrupt the outer membrane by targeting LPS. Our results suggest that microfluidic electrokinetic platforms employing 3DiDEP can be a useful tool for isolating and selecting bacteria based on their LPS glycoforms. Future iterations of these platforms could be leveraged for rapid profiling of pathogens based on their surface LPS structural identity.https://www.frontiersin.org/articles/10.3389/fbioe.2023.991784/fulllipopolysaccharidedielectrophoresislinear electrokineticsouter membrane permeabilitycolistin susceptibility |
| spellingShingle | Qianru Wang Hyungseok Kim Tiffany M. Halvorsen Sijie Chen Christopher S. Hayes Cullen R. Buie Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli Frontiers in Bioengineering and Biotechnology lipopolysaccharide dielectrophoresis linear electrokinetics outer membrane permeability colistin susceptibility |
| title | Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli |
| title_full | Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli |
| title_fullStr | Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli |
| title_full_unstemmed | Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli |
| title_short | Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli |
| title_sort | leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in e coli |
| topic | lipopolysaccharide dielectrophoresis linear electrokinetics outer membrane permeability colistin susceptibility |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.991784/full |
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