Fluorescence-based in situ assay to probe the viability and growth kinetics of surface-adhering and suspended recombinant bacteria
dc.contributor.author
Avalos Vizcarra, Ima
dc.contributor.author
Emge, Philippe
dc.contributor.author
Miermeister, Philipp
dc.contributor.author
Chabria, Mamta
dc.contributor.author
Konradi, Rupert
dc.contributor.author
Vogel, Viola
dc.contributor.author
Möller, Jens
dc.date.accessioned
2019-03-25T14:20:57Z
dc.date.available
2017-06-10T22:01:38Z
dc.date.available
2019-03-25T14:20:57Z
dc.date.issued
2013-12
dc.identifier.issn
1559-4106
dc.identifier.issn
1934-8630
dc.identifier.other
10.1186/1559-4106-8-22
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/72467
dc.identifier.doi
10.3929/ethz-b-000072467
dc.description.abstract
Bacterial adhesion and biofilm growth can cause severe biomaterial-related infections and failure of medical implants. To assess the antifouling properties of engineered coatings, advanced approaches are needed for in situ monitoring of bacterial viability and growth kinetics as the bacteria colonize a surface. Here, we present an optimized protocol for optical real-time quantification of bacterial viability. To stain living bacteria, we replaced the commonly used fluorescent dye SYTO® 9 with endogenously expressed eGFP, as SYTO® 9 inhibited bacterial growth. With the addition of nontoxic concentrations of propidium iodide (PI) to the culture medium, the fraction of live and dead bacteria could be continuously monitored by fluorescence microscopy as demonstrated here using GFP expressing Escherichia coli as model organism. The viability of bacteria was thereby monitored on untreated and bioactive dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAC)-coated glass substrates over several hours. Pre-adsorption of the antimicrobial surfaces with serum proteins, which mimics typical protein adsorption to biomaterial surfaces upon contact with host body fluids, completely blocked the antimicrobial activity of the DMOAC surfaces as we observed the recovery of bacterial growth. Hence, this optimized eGFP/PI viability assay provides a protocol for unperturbed in situ monitoring of bacterial viability and colonization on engineered biomaterial surfaces with single-bacteria sensitivity under physiologically relevant conditions.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
AVS
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/2.0/
dc.subject
Antimicrobial surfaces
en_US
dc.subject
Optical viability monitoring
en_US
dc.subject
Green fluorescent protein (GFP)
en_US
dc.subject
SYTO® 9
en_US
dc.subject
Propidium iodide (PI)
en_US
dc.title
Fluorescence-based in situ assay to probe the viability and growth kinetics of surface-adhering and suspended recombinant bacteria
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 2.0 Generic
dc.date.published
2013-08-21
ethz.journal.title
Biointerphases
ethz.journal.volume
8
en_US
ethz.journal.issue
1
en_US
ethz.pages.start
22
en_US
ethz.size
9 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.scopus
ethz.publication.place
New York, NY
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02540 - Institut für Translationale Medizin / Institute of Translational Medicine::03640 - Vogel, Viola / Vogel, Viola
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02540 - Institut für Translationale Medizin / Institute of Translational Medicine::03640 - Vogel, Viola / Vogel, Viola
ethz.date.deposited
2017-06-10T22:01:59Z
ethz.source
ECIT
ethz.identifier.importid
imp5936510c08ffb17696
ethz.ecitpid
pub:114801
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-08-03T08:38:54Z
ethz.rosetta.lastUpdated
2020-02-15T18:02:45Z
ethz.rosetta.versionExported
true
ethz.COinS
ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.atitle=Fluorescence-based%20in%20situ%20assay%20to%20probe%20the%20viability%20and%20growth%20kinetics%20of%20surface-adhering%20and%20suspended%20recombinant%20bacteria&rft.jtitle=Biointerphases&rft.date=2013-12&rft.volume=8&rft.issue=1&rft.spage=22&rft.issn=1559-4106&1934-8630&rft.au=Avalos%20Vizcarra,%20Ima&Emge,%20Philippe&Miermeister,%20Philipp&Chabria,%20Mamta&Konradi,%20Rupert&rft.genre=article&rft_id=info:doi/10.1186/1559-4106-8-22&
Files in this item
Publication type
-
Journal Article [127708]