Directly use DH5α fdhF+RFP colony to inoculate silicone tube set
with 5ml LB. rest for 12h for biofilm attachment.
Add 50mlLB+Amp and start circulation.
Biofilm with direct inoculation is optimum. Expressed red
florescence but only one small fraction of the biofilm remained after
cutting off the part of the tube.
4 silicone tube sets with biofilm attachement time of 3h, 5h,
7.5h and 10h; 4 bubbling sets.
The silicone tube set with 3h of attachment achieves optimum
biofilm formation. Bubbling sets glass slides are froze in -80鈩for
later observation with laser confocal. Biofilm formed on glass slide is
about 10μm thick.
New sets with silicone tube, a mixture of RFP, CFP and YFP
expressing e.coli. Attachment 3h.
Glass slide with bubbling set with the same mixture of e.coli.
Place two filter film on solid culture medium and placed the mixture of
e.coli on the filter to form gas-solid interface biofilm.
Observation of glass slide and gas-solid biofilm under laser
confocal. Glass slide are observed directly and gas-solid biofilm is
first pressed by a covering slide so that it would stick to it and
peeled off from the filter for observation.
Biofilm on glass slide is about 8.6μm thick. Biofilm formed on gas-solid
interface is too concentrated in bacteria and culture medium that the
florescence of too strong to see any clear structure.