Outlining the strain, growth media and analysis statistics for the different bacteria species used for these experiments.

The growth media was Luria-Bertani (LB) Broth and Mueller Hinton (MH) Broth. We started using LB Broth with E.coli, but switched to MH Broth for the later experiments with P.aeruginosa and S.aureus as we wanted to test with this media as well, given it is recommended for use with AST by EUCAST [44]. The same growth media was used for precultures and preparing pads on the MAP platform. Pad count, microcolony count and bacteria count are reported from a single timepoint around 2.5 hours after imaging was started, meaning each microcolony and cell counted are unique. For the analysis, data from a few consecutive time steps is typically used.