Pseudomonas aeruginosa and Saccharomyces cerevisiae biofilm in flow cells
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Pseudomonas aeruginosa and Saccharomyces cerevisiae biofilm in flow cells. / Weiss Nielsen, Martin; Sternberg, Claus; Molin, Søren; Regenberg, Birgitte.
In: Journal of Visualized Experiments, Vol. 47, 2011, p. pii:2383.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Pseudomonas aeruginosa and Saccharomyces cerevisiae biofilm in flow cells
AU - Weiss Nielsen, Martin
AU - Sternberg, Claus
AU - Molin, Søren
AU - Regenberg, Birgitte
PY - 2011
Y1 - 2011
N2 - Many microbial cells have the ability to form sessile microbial communities defined as biofilms that have altered physiological and pathological properties compared to free living microorganisms. Biofilms in nature are often difficult to investigate and reside under poorly defined conditions(1). Using a transparent substratum it is possible to device a system where simple biofilms can be examined in a non-destructive way in real-time: here we demonstrate the assembly and operation of a flow cell model system, for in vitro 3D studies of microbial biofilms generating high reproducibility under well-defined conditions(2,3). The system consists of a flow cell that serves as growth chamber for the biofilm. The flow cell is supplied with nutrients and oxygen from a medium flask via a peristaltic pump and spent medium is collected in a waste container. This construction of the flow system allows a continuous supply of nutrients and administration of e.g. antibiotics with minimal disturbance of the cells grown in the flow chamber. Moreover, the flow conditions within the flow cell allow studies of biofilm exposed to shear stress. A bubble trapping device confines air bubbles from the tubing which otherwise could disrupt the biofilm structure in the flow cell. The flow cell system is compatible with Confocal Laser Scanning Microscopy (CLSM) and can thereby provide highly detailed 3D information about developing microbial biofilms. Cells in the biofilm can be labeled with fluorescent probes or proteins compatible with CLSM analysis. This enables online visualization and allows investigation of niches in the developing biofilm. Microbial interrelationship, investigation of antimicrobial agents or the expression of specific genes, are of the many experimental setups that can be investigated in the flow cell system.
AB - Many microbial cells have the ability to form sessile microbial communities defined as biofilms that have altered physiological and pathological properties compared to free living microorganisms. Biofilms in nature are often difficult to investigate and reside under poorly defined conditions(1). Using a transparent substratum it is possible to device a system where simple biofilms can be examined in a non-destructive way in real-time: here we demonstrate the assembly and operation of a flow cell model system, for in vitro 3D studies of microbial biofilms generating high reproducibility under well-defined conditions(2,3). The system consists of a flow cell that serves as growth chamber for the biofilm. The flow cell is supplied with nutrients and oxygen from a medium flask via a peristaltic pump and spent medium is collected in a waste container. This construction of the flow system allows a continuous supply of nutrients and administration of e.g. antibiotics with minimal disturbance of the cells grown in the flow chamber. Moreover, the flow conditions within the flow cell allow studies of biofilm exposed to shear stress. A bubble trapping device confines air bubbles from the tubing which otherwise could disrupt the biofilm structure in the flow cell. The flow cell system is compatible with Confocal Laser Scanning Microscopy (CLSM) and can thereby provide highly detailed 3D information about developing microbial biofilms. Cells in the biofilm can be labeled with fluorescent probes or proteins compatible with CLSM analysis. This enables online visualization and allows investigation of niches in the developing biofilm. Microbial interrelationship, investigation of antimicrobial agents or the expression of specific genes, are of the many experimental setups that can be investigated in the flow cell system.
KW - Biofilms
KW - Flow Cytometry
KW - Pseudomonas aeruginosa
KW - Saccharomyces cerevisiae
U2 - 10.3791/2383
DO - 10.3791/2383
M3 - Journal article
C2 - 21304454
VL - 47
SP - pii:2383
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
ER -
ID: 35231687