Two-dimensional and three-dimensional dynamic imaging of live biofilms in a microchannel by time-of-flight secondary ion mass

A vacuum compatible microfluidic reactor, SALVI (System for Analysis at the Liquid Vacuum Interface) was employed for in situ chemical imaging of live biofilms using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Depth profiling by sputtering materials in sequential layers resulted in live biofilm spatial chemical mapping. 2D images were reconstructed to report the first 3D images of hydrated biofilm elucidating spatial and chemical heterogeneity. 2D image principal component analysis (PCA) was conducted among biofilms at different locations in the microchannel. Our approach directly visualized spatial and chemical heterogeneity within the living biofilm by dynamic liquid ToF-SIMS.