Confocal microscopy images for: Surface remodeling and inversion of cell-matrix interactions underlie community recognition and dispersal in Vibrio cholerae biofilms
Data files
Dec 04, 2024 version files 1.59 GB
-
Dataset_1_Fig1b_Timecourse.nd2
573.38 MB
-
Dataset_2_Fig4a_Timecourse.nd2
424.59 MB
-
Dataset_3_Fig3c_VPS-Bap1.nd2
592.02 MB
-
README.md
854 B
Abstract
Biofilms are ubiquitous surface-associated bacterial communities embedded in an extracellular matrix. It is commonly assumed that biofilm cells are glued together by the matrix; however, how the specific biochemistry of matrix components affects the cell-matrix interactions and how these interactions vary during biofilm growth remain unclear. Here, we investigate cell-matrix interactions in Vibrio cholerae, the causative agent of cholera. We combine genetics, microscopy, simulations, and biochemical analyses to show that V. cholerae cells are not attracted to the main matrix component (Vibrio polysaccharide, VPS), but can be attached to each other and to the VPS network through crosslinks formed by protein Bap1. Downregulation of VPS production and surface trimming by polysaccharide lyase RbmB cause surface remodeling as biofilms age, shifting the nature of cell-matrix interactions from attractive to repulsive and facilitating cell dispersal as aggregated groups. Our results shed light into the dynamics of diverse cell-matrix interactions as drivers of biofilm development.
README: Confocal microscopy images for: Surface remodeling and inversion of cell-matrix interactions underlie community recognition and dispersal in Vibrio cholerae biofilms
https://doi.org/10.5061/dryad.zcrjdfnph
Description of the data and file structure
Confocal microscopy images for Vibrio cholerae biofilm for: Surface remodeling and inversion of cell-matrix interactions underlie community recognition and dispersal in Vibrio cholerae biofilms
Files and variables
File: Dataset_1_Fig1b_Timecourse.nd2
Description: Raw data for Figure 1b.
File: Dataset_2_Fig4a_Timecourse.nd2
Description: Raw data for Figure 4a.
File: Dataset_3_Fig3c_VPS-Bap1.nd2
Description: Raw data for Figure 3c.
Code/software
FIJI 2.16.0 can be used to open the nd2 files.
Methods
All datasets were acquired using a Yokogawa CSU-W1 spinning-disk confocal scanning unit mounted on a Nikon Ti2-E microscope body, using the Nikon perfect focus system and images were acquired using Nikon Elements 5.20. For time course imaging, cells were incubated in a Tokai-Hit stage-top incubator at a temperature of 25°C. Provided here are raw data in nd2 format, which can be opened with FIJI.
Dataset 1: For this data, a 60× water immersion objective (numerical aperture = 1.20) was used. FM 4-64 at 2 µg/mL was used to stain cell membranes and imaged at 561 nm. Time difference between images is 30 minutes.
Dataset 2: For this data, a 100× silicon oil immersion objective (Lambda S 100XC Sil, numerical aperture = 1.35) was used. FM 4-64 at 2 µg/mL was used to stain cell membranes and imaged at 561 nm. Cells consistutively express SCFP3A and were imaged at 445 nm, although the signals were weak. VPS was stained with wheat germ agglutinin (WGA) conjugated to Oregon Green (4 µg/mL). Time difference between images is 30 minutes.
Dataset 3: For this data, a 100× silicon oil immersion objective (Lambda S 100XC Sil, numerical aperture = 1.35) was used. A z-step size of 0.13 μm was used. FM 4-64 at 2 µg/mL was used to stain cell membranes and imaged at 561 nm. VPS was stained with wheat germ agglutinin (WGA) conjugated to AlexaFluor647 (4 µg/mL). Bap1-GFP was imaged at 488 nm.