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Trouble with imaging of Pseudomonas aeruginosa

Mark nospam at nspam.com
Fri Jul 30 07:45:57 EST 2004

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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