Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion
Abstract Bacterial biofilms are complex cell communities within a self-produced extracellular matrix, crucial in various fields but challenging to analyze in 3D. We developed a “biofilm-in-capillary” growth method compatible with full-rotation soft X-ray tomography, enabling high-resolution 3D imagi...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-02-01
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| Series: | npj Biofilms and Microbiomes |
| Online Access: | https://doi.org/10.1038/s41522-025-00659-0 |
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| author | Anthoula Chatzimpinou Anne Diehl A. Tobias Harhoff Kristina Driller Bieke Vanslembrouck Jian-Hua Chen Kristaps Kairišs Valentina Loconte Mark A. Le Gros Carolyn Larabell Kürşad Turgay Hartmut Oschkinat Venera Weinhardt |
| author_facet | Anthoula Chatzimpinou Anne Diehl A. Tobias Harhoff Kristina Driller Bieke Vanslembrouck Jian-Hua Chen Kristaps Kairišs Valentina Loconte Mark A. Le Gros Carolyn Larabell Kürşad Turgay Hartmut Oschkinat Venera Weinhardt |
| author_sort | Anthoula Chatzimpinou |
| collection | DOAJ |
| description | Abstract Bacterial biofilms are complex cell communities within a self-produced extracellular matrix, crucial in various fields but challenging to analyze in 3D. We developed a “biofilm-in-capillary” growth method compatible with full-rotation soft X-ray tomography, enabling high-resolution 3D imaging of bacterial cells and their matrix during biofilm formation. This approach offers 50 nm isotropic spatial resolution, rapid imaging, and quantitative native analysis of biofilm structure. Using Bacillus subtilis biofilms, we detected coherent alignment and chaining of wild-type cells towards the oxygen-rich capillary tip. In contrast, the ΔtasA genetic knock-out showed a loss of cellular orientation and changes in the extracellular matrix. Adding TasA protein to the ΔtasA strain restored matrix density and led to cell assembly compaction, but without the chaining observed in wild-type biofilms. This scalable and transferable approach opens new avenues for examining biofilm structure and function across various species, including mixed biofilms, and response to genetic and environmental factors. |
| format | Article |
| id | doaj-art-15954240ca444ab2b1713e35a345b641 |
| institution | Kabale University |
| issn | 2055-5008 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Biofilms and Microbiomes |
| spelling | doaj-art-15954240ca444ab2b1713e35a345b6412025-02-09T12:15:21ZengNature Portfolionpj Biofilms and Microbiomes2055-50082025-02-0111111010.1038/s41522-025-00659-0Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletionAnthoula Chatzimpinou0Anne Diehl1A. Tobias Harhoff2Kristina Driller3Bieke Vanslembrouck4Jian-Hua Chen5Kristaps Kairišs6Valentina Loconte7Mark A. Le Gros8Carolyn Larabell9Kürşad Turgay10Hartmut Oschkinat11Venera Weinhardt12Centre for Organismal Studies, Heidelberg UniversityLeibniz-Forschungsinstitut für Molekulare PharmakologieCentre for Organismal Studies, Heidelberg UniversityMax Planck Unit for the Science of Pathogens, Leibniz UniversitätMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National LaboratoryMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National LaboratoryCentre for Organismal Studies, Heidelberg UniversityMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National LaboratoryMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National LaboratoryMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National LaboratoryMax Planck Unit for the Science of Pathogens, Leibniz UniversitätLeibniz-Forschungsinstitut für Molekulare PharmakologieCentre for Organismal Studies, Heidelberg UniversityAbstract Bacterial biofilms are complex cell communities within a self-produced extracellular matrix, crucial in various fields but challenging to analyze in 3D. We developed a “biofilm-in-capillary” growth method compatible with full-rotation soft X-ray tomography, enabling high-resolution 3D imaging of bacterial cells and their matrix during biofilm formation. This approach offers 50 nm isotropic spatial resolution, rapid imaging, and quantitative native analysis of biofilm structure. Using Bacillus subtilis biofilms, we detected coherent alignment and chaining of wild-type cells towards the oxygen-rich capillary tip. In contrast, the ΔtasA genetic knock-out showed a loss of cellular orientation and changes in the extracellular matrix. Adding TasA protein to the ΔtasA strain restored matrix density and led to cell assembly compaction, but without the chaining observed in wild-type biofilms. This scalable and transferable approach opens new avenues for examining biofilm structure and function across various species, including mixed biofilms, and response to genetic and environmental factors.https://doi.org/10.1038/s41522-025-00659-0 |
| spellingShingle | Anthoula Chatzimpinou Anne Diehl A. Tobias Harhoff Kristina Driller Bieke Vanslembrouck Jian-Hua Chen Kristaps Kairišs Valentina Loconte Mark A. Le Gros Carolyn Larabell Kürşad Turgay Hartmut Oschkinat Venera Weinhardt Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion npj Biofilms and Microbiomes |
| title | Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion |
| title_full | Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion |
| title_fullStr | Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion |
| title_full_unstemmed | Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion |
| title_short | Soft X-ray tomography reveals variations in B. subtilis biofilm structure upon tasA deletion |
| title_sort | soft x ray tomography reveals variations in b subtilis biofilm structure upon tasa deletion |
| url | https://doi.org/10.1038/s41522-025-00659-0 |
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