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Biofilm
techniques

Essential elements to understanding the
function of biofilms
 determining factors influencing the
processes of:
- bacterial adhesion and biofilm formation
- spatial arrangement of biofilms including
distribution and composition of
microorganisms within the matrix

 characterizing the properties of the
matrix

“in vitro“ systems
Steady-state systems
(microtiter plates, TSP, CBD
...
)

“in vivo“ systems
infection models

Steady-state systems
Microtiter plates

Calgary Biofilm Device

MBEC determination

Continuous flow systems

Flow cells system

Rotating disc reactor

CDC reactor

Robbins Device

Biomass quantification
Cristaviolet assay (Christensen, 1985)
Basic dye that bound to negative charged molecules present on cell
surface and in the matrix
...


ALAMAR BLU

FDA

The oxidated form
(resazurin) is dark blue the
reduced form (resorufin)
is violet and fluorescent
...


Matrix quantification
 DMMB (1,9-dimethyl methylene blue)
DMMB forms an insoluble complexation product with sulphated polysaccharides
in the biofilm matrix
...

- Propidio iodide → fluorogenic dye
(red) which doesn’t diffuse
passively through cellular
membranes
...
(pathogenesis,
tissue damage, inflammation, immune response)
 Evaluate the level of virulence of the infecting bacteria
 Evalute protocolos to prevent biofilm-related infections
 Evaluate the effect of treatment toward biofilm-related infections

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