Dense initial population cultures can be used to measure lag time. Lag time is defined as the delay that a culture demonstrates from the time it is freshly inoculated to the time that it achieves its minimal doubling time. The spline model was used to measure lag for a representative culture printed at (a) lower dilution (1:4) and (b) higher dilution (1:2000). In panel (a), lag time can be directly observed, because the spot is detectable (average pixel intensity > 1) at time = 0. In contrast, in panel (b) cultures have passed through lag phase and exhibit MSR by the time spot intensities reach the threshold of detection.