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Biofilm is a multi-cellular bacteria community, which is
prevalent in diverse natural and artificial settings. It is best known
for its robust adaptability and strong resistance to environmental
stress. We intended to utilize and modify this natural multicellular
structure to develop a new kind of extracellular scaffold, constructing
a stratified protein expression system using the naturally formed graded
oxygen concentration within the bacteria biofilm.
Biofilm formation process can be roughly divided into four
stages: bacteria attachment, micro colony formation, biofilm maturation
and biofilm dispersion. The best sample for the observation is the
matured biofilm, which, according to previous studies, was formed after
48 hours after inoculation with only exception in bubbling method
(Bandara HM et al. 2009; Klausen M. et al. 2003.).
Based on the references (Almeida C. et al. 2011; Rani SA et al. 2007), we’ve developed three different biofilm formation methods: Bubbling method, Rubber tube method and cell culture plate method. The biofilm formed in the two former ways were observed through cryo-sectioning coupled with normal fluorescence microscopy while confocal laser scanning microscope was employed to obtain the biofilm images of the third method. In addition to the above, we’ve tested different material’s applicability in biofilm formation experiments and introduced autoinducer AI-2 as an accelerator molecule.