High Content Imaging: Using confocal microscopy to collect 3D and 4D biofilm growth data

A Pseudomonas fluorescens biofilm was grown in minimal media for 24 hours and stained with BacLight Viability Stain (Invitrogen). The live biofilm was imaged on a Nikon TE2000U inverted scope with Nikon Live-Scan Swept-Field Confocal with a 40X ELWD objective corrected for the 180μm thickness of the bottom of the BioFlux Plate. Data were collected using a Photometrics Cascade camera and NIS-Elements software in 1.5 μm slices over a Z-distance of 70μm. This distance covers the complete height of the BioFlux microfludic channel in the viewing window region of the plate.