Monitoring of biofilms grown on differentially structured metallic surfaces using confocal laser scanning microscopy
Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a h...
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| Published in | Engineering in life sciences Vol. 19; no. 7; pp. 513 - 521 |
|---|---|
| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Germany
John Wiley and Sons Inc
01.07.2019
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1618-0240 1618-2863 |
| DOI | 10.1002/elsc.201800176 |
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| Abstract | Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)‐expressing Escherichia coli strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of E. coli GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP‐expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm. |
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| AbstractList | Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)‐expressing Escherichia coli strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of E. coli GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP‐expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm. Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)-expressing Escherichia coli strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of E. coli GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP-expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm.Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)-expressing Escherichia coli strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of E. coli GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP-expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm. Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)-expressing strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP-expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm. Imaging of biofilms on opaque surfaces is a challenge presented to researchers especially considering pathogenic bacteria, as those typically grow on living tissue, such as mucosa and bone. However, they can also grow on surfaces used in industrial applications such as food production, acting as a hindrance to the process. Thus, it is important to understand bacteria better in the environment they actually have relevance in. Stainless steel and titanium substrata were line structured and dotted surface topographies for titanium substrata were prepared to analyze their effects on biofilm formation of a constitutively green fluorescent protein (GFP)‐expressing Escherichia coli strain. The strain was batch cultivated in a custom built flow cell initially for 18 h, followed by continuous cultivation for 6 h. Confocal laser scanning microscopy (CLSM) was used to determine the biofilm topography. Biofilm growth of E. coli GFPmut2 was not affected by the type of metal substrate used; rather, attachment and growth were influenced by variable shapes of the microstructured titanium surfaces. In this work, biofilm cultivation in flow cells was coupled with the most widely used biofilm analytical technique (CLSM) to study the time course of growth of a GFP‐expressing biofilm on metallic surfaces without intermittent sampling or disturbing the natural development of the biofilm. |
| Author | Müller‐Renno, Christine Mukherjee, Joydeep Schlegel, Christin Mitra, Sayani Chodorski, Jonas Huttenlochner, Katharina Ziegler, Christiane Ulber, Roland Kleine, Daniel |
| AuthorAffiliation | 3 Department of Physics and Research Center OPTIMAS TU Kaiserslautern Kaiserslautern Germany 1 Institute of Bioprocess Engineering TU Kaiserslautern Kaiserslautern Germany 2 School of Environmental Studies Jadavpur University Kolkata India |
| AuthorAffiliation_xml | – name: 2 School of Environmental Studies Jadavpur University Kolkata India – name: 1 Institute of Bioprocess Engineering TU Kaiserslautern Kaiserslautern Germany – name: 3 Department of Physics and Research Center OPTIMAS TU Kaiserslautern Kaiserslautern Germany |
| Author_xml | – sequence: 1 givenname: Daniel surname: Kleine fullname: Kleine, Daniel organization: TU Kaiserslautern – sequence: 2 givenname: Jonas surname: Chodorski fullname: Chodorski, Jonas organization: TU Kaiserslautern – sequence: 3 givenname: Sayani surname: Mitra fullname: Mitra, Sayani organization: Jadavpur University – sequence: 4 givenname: Christin surname: Schlegel fullname: Schlegel, Christin organization: TU Kaiserslautern – sequence: 5 givenname: Katharina surname: Huttenlochner fullname: Huttenlochner, Katharina organization: TU Kaiserslautern – sequence: 6 givenname: Christine surname: Müller‐Renno fullname: Müller‐Renno, Christine organization: TU Kaiserslautern – sequence: 7 givenname: Joydeep surname: Mukherjee fullname: Mukherjee, Joydeep organization: Jadavpur University – sequence: 8 givenname: Christiane surname: Ziegler fullname: Ziegler, Christiane organization: TU Kaiserslautern – sequence: 9 givenname: Roland orcidid: 0000-0002-7674-0967 surname: Ulber fullname: Ulber, Roland email: ulber@mv.uni-kl.de organization: TU Kaiserslautern |
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| Title | Monitoring of biofilms grown on differentially structured metallic surfaces using confocal laser scanning microscopy |
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