Trouble with imaging of Pseudomonas aeruginosa

Lewandowski, Zbigniew zl at erc.montana.edu
Mon Aug 16 10:19:30 EST 2004


Ashley: You are not the only one who has doubts about what the available
software packages are actually quantifying from the images of biofilms. I
suggest that you get a copy of our recent paper: Beyenal H., Lewandowski Z.,
Harkin G. (2004). Quantifying biofilm structure: facts and fiction.
Biofouling. 20: 1-23.It addresses many of your questions, and also specifies
what ISA does and does not. A new version of ISA, which can quantify 3D
biofilm structure, will be presented at Biofilms 2004 in Las Vegas.
Zbigniew Lewandowski


 >  Zbigniew Lewandowski
 >  Professor
 >  Department of Civil Engineering
 >  and Center for Biofilm Engineering
 >  Montana State University
 >  EPS Building, Room 310
 >  Bozeman, MT 59717
 >  Tel: (406)-994-5915   Fax:(406)-994-6098
 >  ZL at erc.montana.edu
 >
 >

-----Original Message-----
From: owner-biofilms at hgmp.mrc.ac.uk [mailto:owner-biofilms at hgmp.mrc.ac.uk]
On Behalf Of nospam at nspam.com
Sent: Friday, July 30, 2004 6:46 AM
To: biofilms at net.bio.net
Subject: Re: Trouble with imaging of Pseudomonas aeruginosa

Ashley Won wrote:
 >Hi all,
 >I'm a postgraduate student working on imaging of Pseudomonas
 >aeruginosa biofilms.
 >After much reading, I came across 2 image analysis programs: COMSTAT
 >and ISA. From my understanding of these 2 programs, neither allows
 >quantification of biofilms using light microscopy. I do not have
 >fluorescent-labeled Pseudomonas aeruginosa thus is not able to utilize
 >COMSTAT. Unfortunately, apart from quantification of structural
 >parameters, ISA does not allow quantification of the whole biofilm.
 >Does anyone know of a program that can allow 3D analysis and
 >quantification of biofilm images taken using light microscopy?
 >I intended to label Pseudomonas aeruginosa with GFP. However, I lack
 >the means of transforming the plasmid into the bacteria since I do not
 >have appropriate conjugative E.coli strain S17-1. Is there any other
 >way to make the bacteria fluoresce throughout the whole experiment?
 >Please help! I'm at a loss of what can I do next.
 >---

Have you looked for S17.1 or another siimlar strain (eg SM10) at ATCC?
Alternatively, if you already have a conjugative (or mobilisable) plasmid
with GFP, you could try a tripartite mating. Along with the P.aeruginosa
and E. coli (GFP) strains, just include an E. coli strain possessing a
conjugative plasmid such as pRK2013 in the mating. The pRK2013 should
provide the mob functions almost as well as the S17.1 could. Afterall,
S17.1 just has RP4 integrated to do that job.
---



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