Finding the comfort zone: Online-monitoring of electroactive bacteria colonising electrode surfaces with different chemical properties

2020 ◽  
Author(s):  
Hanna Frühauf ◽  
Markus Stöckl ◽  
Dirk Holtmann

<p>Mechanisms of electron transfer vary greatly within the diverse group of electroactive microorganisms and so does the need to attach to the electrode surface, e.g. by forming a biofilm.</p> <p>Electrochemical impedance spectroscopy (EIS) and confocal laser scanning microscopy (CLSM) are well established methods to monitor cell attachment to an electrode surface and have therefore been combined in a flow cell as a screening system. The flow cell, equipped with a transparent indium tin oxide working electrode (ITO WE), allows monitoring of attachment processes in real time with minimal needs for additional biofilm preparation. In preliminary experiments the flow cell was successfully used as microbial fuel cell (MFC) with a potential of +0.4 V vs. Ag/AgCl using <em>Shewanella oneidensis</em> as electroactive model organism. [1]</p> <p>Commonly, graphite-based electrode materials are used in bioelectrochemical systems due to their low costs and high conductivity. However, the hydrophobic and negatively charged surface is not yet optimal for microbial attachment. There are numerous attempts on electrode surface engineering in order to overcome this problem. In the majority of studies the biofilm analysis and evaluation of the attachment takes place at the end of the experiment, neglecting the impacts of the chemical surface properties and initial electrode conditioning during the very beginning of biofilm formation.</p> <p>To investigate initial attachment and biofilm formation in real-time, the transparent ITO-electrode is coated with polyelectrolytes differing in hydrophobicity and polarity to evaluate their effects on the initial surface colonisation by different electroactive microorganisms. Combining CLSM and EIS, both, surface coverage and electrochemical interaction of electrode-associated bacteria can be assessed.</p> <p>With this we aim to understand and ease initial steps of biofilm formation to improve efficiency of bioelectrochemical applications, e.g. with regards to start-up time.</p> <p> </p> <p>[1] Stöckl, M., Schlegel, C., Sydow, A., Holtmann, D., Ulber, R., & Mangold, K. M. (2016). Membrane separated flow cell for parallelized electrochemical impedance spectroscopy and confocal laser scanning microscopy to characterize electro-active microorganisms. <em>Electrochimica Acta</em>, 220, 444-452.</p>

2003 ◽  
Vol 47 (5) ◽  
pp. 139-142 ◽  
Author(s):  
A.C. Rodrigues ◽  
S. Wuertz ◽  
A.G. Brito ◽  
L.F. Melo

Confocal laser scanning microscopy was used to monitor the colonization pattern of the gfp-labeled derivative strain of Pseudomonas putida ATCC 17514 on fluorene and phenanthrene crystals. The in situ experiments showed that P. putida tends to grow directly on phenanthrene, forming a biofilm on accessible crystalline surfaces. On the other hand, no significant biofilm formation was observed in the presence of fluorene. The results obtained showed that substrate properties affected bacterial strategy regarding uptake.


2021 ◽  
Author(s):  
Shayesteh Beladi-Behbahani ◽  
Sarah M. Helms ◽  
John D. DesJardins ◽  
Marian S. Kennedy ◽  
Terri Bruce ◽  
...  

Abstract In order to assess and compare the antibacterial property of implants surfaces, a standard method is needed to quantify bacterial load. This study evaluated the effectiveness of three quantifying methods, namely, (I) crystal violet staining analysis, (II) ultrasound detachment with viable cell counts, and (III) confocal laser scanning microscopy for characterizing S. aureus Seattle 1945 (ATCC 25923) biofilm on metallic coupons. The accuracy of the results, time for completion, and ease of use of methods were compared. The crystal violet method is relatively faster and more straightforward for analyzing biofilm formation. However, the accuracy of the confocal laser scanning microscopy method is found to be considerably higher than that of the other methods. Confocal laser scanning microscopy method is considered to be more time-consuming for data collection and analysis and costlier. The ultrasound detachment followed by viable cell count of recovered cells is recommended for biofilm quantification analysis on orthopedic materials when there is a large number of samples (more than ten samples). This info could provide guidelines that would facilitate the selection of suitable method for quantifying biofilm formation on orthopedic implants based on investigators’ consideration on method accessibility, assay cost, assay time, and complexity of method.


2007 ◽  
Vol 189 (22) ◽  
pp. 8353-8356 ◽  
Author(s):  
Luyan Ma ◽  
Haiping Lu ◽  
April Sprinkle ◽  
Matthew R. Parsek ◽  
Daniel J. Wozniak

ABSTRACT The Pseudomonas aeruginosa polysaccharide synthesis locus (psl) is predicted to encode an exopolysaccharide which is critical for biofilm formation. Here we used chemical composition analyses and mannose- or galactose-specific lectin staining, followed by confocal laser scanning microscopy and electron microscopy, to show that Psl is a galactose-rich and mannose-rich exopolysaccharide.


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