In order to enable better diffusion of the tested compound into the agar surface, the plates can stay at low temperature
for a few hours before incubation. This method is a combination
of contact and direct bioautography, because the antimicrobial
compounds are transferred from the chromatogram to the agar
medium, as in a contact method, but the agar layer remains onto
the chromatogram surface during the incubation and visualization,
as in direct bioautography.