Journal: Nature Communications

Article Title: The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction

doi: 10.1038/ncomms11951

Figure Lengend Snippet: ( a ) Representative image of a P. aeruginosa biofilm grown in a drip flow reactor (DFR). The flow of nutrients is depicted by the topmost arrows ranging in colour from blue to brown. Blue boxes denote the approximate regions from which 12-μm-thick sections were obtained for MALDI IMS analysis. ( b , c ) MALDI IMS signals with differential biofilm localization found primarily in portions of the biomass presumed to be nutrient-replete. ( d , e ) MALDI IMS signals primarily localized to portions of the biomass predicted to be nutrient-deplete. ( f ) Overlay of signals shown in b – e highlighting sublocalization of signals within the predicted nutrient-replete and nutrient-deplete niches with zoomed insets highlighted. Scale bar, 3 mm (inset scale bars, 1 mm).

Article Snippet: Biofilms were grown in a Drip Flow Biofilm Reactor (DFR) (BioSurface Technologies, Bozeman, MT) similar to previously described methods using glass microscope slides as the growth surface.

Techniques:

Journal: Nature Communications

Article Title: The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction

doi: 10.1038/ncomms11951

Figure Lengend Snippet: m/z values and associated protein designations are included adjacent to MALDI IMS images of the biofilm section proximal to the nutrient pore highlighted in . These identifications derive from signals that were reproducibly detected during the analysis of 15 replicate DFR biofilms. Scale bar, 3 mm.

Article Snippet: Biofilms were grown in a Drip Flow Biofilm Reactor (DFR) (BioSurface Technologies, Bozeman, MT) similar to previously described methods using glass microscope slides as the growth surface.

Techniques:

Journal: Nature Communications

Article Title: The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction

doi: 10.1038/ncomms11951

Figure Lengend Snippet: ( a ) LA-ICP IMS analysis of a biofilm section containing a defined ‘central channel' and ‘nutrient-deplete edge'. Scale bar, 2 mm. ( b ) ICP-MS quantification of metal levels within distinct portions of the biofilm. Error bars represent s.d. of data derived from triplicate biofilms. ‘*' designates P <0.05, ‘**' designates P <0.002 as determined by a Student's t -test. ( c ) MALDI IMS analysis highlights the presence of differential protein distribution patterns that follow similar trends to the metal localization patterns. Scale bar, 3 mm. NS, not significant.

Article Snippet: Biofilms were grown in a Drip Flow Biofilm Reactor (DFR) (BioSurface Technologies, Bozeman, MT) similar to previously described methods using glass microscope slides as the growth surface.

Techniques: Derivative Assay

Journal: Nature Communications

Article Title: The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction

doi: 10.1038/ncomms11951

Figure Lengend Snippet: HcnB, PhzS and PqsB are components of various anti-staphylococcal biosynthetic pathways that were found in lower abundance in the biofilm edge. While expression of the corresponding genes has been shown to be both Fe and quorum sensing (QS)-regulated, the proteins' distribution profiles did not correlate with those of known Fe-responsive proteins such as FpvA and SdhA or proteins encoded by known QS-responsive genes such as LasA and RmlC. Error bars are s.e.m. derived from biological triplicate samples processed in four technical replicates per biofilm. ‘*' denotes P <0.05, ‘**' denotes P <0.02 as determined by a Student's t -test. NS, not significant.

Article Snippet: Biofilms were grown in a Drip Flow Biofilm Reactor (DFR) (BioSurface Technologies, Bozeman, MT) similar to previously described methods using glass microscope slides as the growth surface.

Techniques: Expressing, Derivative Assay