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Non-invasive characterization of electrochemically active microbial biofilms using confocal Raman microscopy

Bernardino Virdis, Falk Harnisch, Damien J Batstone, Korneel Rabaey UGent and Bogdan C Donose (2012) ENERGY & ENVIRONMENTAL SCIENCE. 5(5). p.7017-7024
abstract
Electrochemically active biofilms rely on microorganisms capable of extracellular electron transfer. Such biofilms are involved in the dissimilatory reduction of metal oxides in natural environments as well as electricity driving and driven processes at the electrodes of microbial bioelectrochemical systems. In this work we present the application of confocal Raman microscopy (CRM) as a non-invasive, label-free, and in vivo characterization method of acetate oxidizing anodic biofilms, grown from primary wastewater inoculum and dominated by Geobacter species (>85% of sequences analysed using pyrotag sequencing). Using the resonance Raman effect of the heme protein cytochrome c (Cyt c)-an ubiquitous component of extracellular electron transfer reactions-it was possible to collect characteristic spectral information of electrochemically active biofilms at pixel integration times of 0.2 s and an excitation wavelength of 532 nm. This allowed monitoring of biofilm development at different growth stages, without impacting its structural or metabolic activity. Furthermore, we demonstrate the possibility of non-invasive investigation of the spatial redox electrochemistry (up to a compositional level) of electrochemically active biofilms, as we observed significant changes in the vibrational properties of Cyt c resulting from shifts in the anodic potential between different redox conditions. Compared to conventional methods requiring destructive sample manipulation and fixation, the proposed approach based on CRM allows the non-invasive analysis of microbial aggregates with minimal sample preparation or prior knowledge of the sample.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
COMMUNITIES, SURFACE, ANODE BIOFILM, GEOBACTER-SULFURREDUCENS, BACTERIAL NANOWIRES, IN-SITU, FUEL-CELLS, RESONANCE RAMAN, SPECTROSCOPY, EXTRACELLULAR ELECTRON-TRANSFER
journal title
ENERGY & ENVIRONMENTAL SCIENCE
Energy Environ. Sci.
volume
5
issue
5
pages
7017 - 7024
Web of Science type
Article
Web of Science id
000303251500043
JCR category
ENVIRONMENTAL SCIENCES
JCR impact factor
11.653 (2012)
JCR rank
2/209 (2012)
JCR quartile
1 (2012)
ISSN
1754-5692
DOI
10.1039/c2ee03374g
project
Biotechnology for a sustainable economy (Bio-Economy)
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2109586
handle
http://hdl.handle.net/1854/LU-2109586
date created
2012-05-21 11:13:42
date last changed
2014-05-26 10:03:08
@article{2109586,
  abstract     = {Electrochemically active biofilms rely on microorganisms capable of extracellular electron transfer. Such biofilms are involved in the dissimilatory reduction of metal oxides in natural environments as well as electricity driving and driven processes at the electrodes of microbial bioelectrochemical systems. In this work we present the application of confocal Raman microscopy (CRM) as a non-invasive, label-free, and in vivo characterization method of acetate oxidizing anodic biofilms, grown from primary wastewater inoculum and dominated by Geobacter species ({\textrangle}85\% of sequences analysed using pyrotag sequencing). Using the resonance Raman effect of the heme protein cytochrome c (Cyt c)-an ubiquitous component of extracellular electron transfer reactions-it was possible to collect characteristic spectral information of electrochemically active biofilms at pixel integration times of 0.2 s and an excitation wavelength of 532 nm. This allowed monitoring of biofilm development at different growth stages, without impacting its structural or metabolic activity. Furthermore, we demonstrate the possibility of non-invasive investigation of the spatial redox electrochemistry (up to a compositional level) of electrochemically active biofilms, as we observed significant changes in the vibrational properties of Cyt c resulting from shifts in the anodic potential between different redox conditions. Compared to conventional methods requiring destructive sample manipulation and fixation, the proposed approach based on CRM allows the non-invasive analysis of microbial aggregates with minimal sample preparation or prior knowledge of the sample.},
  author       = {Virdis, Bernardino and Harnisch, Falk and Batstone, Damien J and Rabaey, Korneel and Donose, Bogdan C},
  issn         = {1754-5692},
  journal      = {ENERGY \& ENVIRONMENTAL SCIENCE},
  keyword      = {COMMUNITIES,SURFACE,ANODE BIOFILM,GEOBACTER-SULFURREDUCENS,BACTERIAL NANOWIRES,IN-SITU,FUEL-CELLS,RESONANCE RAMAN,SPECTROSCOPY,EXTRACELLULAR ELECTRON-TRANSFER},
  language     = {eng},
  number       = {5},
  pages        = {7017--7024},
  title        = {Non-invasive characterization of electrochemically active microbial biofilms using confocal Raman microscopy},
  url          = {http://dx.doi.org/10.1039/c2ee03374g},
  volume       = {5},
  year         = {2012},
}

Chicago
Virdis, Bernardino, Falk Harnisch, Damien J Batstone, Korneel Rabaey, and Bogdan C Donose. 2012. “Non-invasive Characterization of Electrochemically Active Microbial Biofilms Using Confocal Raman Microscopy.” Energy & Environmental Science 5 (5): 7017–7024.
APA
Virdis, Bernardino, Harnisch, F., Batstone, D. J., Rabaey, K., & Donose, B. C. (2012). Non-invasive characterization of electrochemically active microbial biofilms using confocal Raman microscopy. ENERGY & ENVIRONMENTAL SCIENCE, 5(5), 7017–7024.
Vancouver
1.
Virdis B, Harnisch F, Batstone DJ, Rabaey K, Donose BC. Non-invasive characterization of electrochemically active microbial biofilms using confocal Raman microscopy. ENERGY & ENVIRONMENTAL SCIENCE. 2012;5(5):7017–24.
MLA
Virdis, Bernardino, Falk Harnisch, Damien J Batstone, et al. “Non-invasive Characterization of Electrochemically Active Microbial Biofilms Using Confocal Raman Microscopy.” ENERGY & ENVIRONMENTAL SCIENCE 5.5 (2012): 7017–7024. Print.