This work involves only data processing software and does not involve data to publish. Data are acquired by the users of this software by the following methods. Two methods can be used to acquire images of agar plates with bacterial colonies grown. In the first, plates were placed on a flatbed scanner, covered by soft gloss paper (HP LaserJet soft gloss paper), and 8-bit grayscale images of the plates were obtained at 300 dpi (dots per inch; 118 dots per cm), 600 dpi (236 dots per cm), and 1,200 dpi (472 dots per cm). In the second approach, images are acquired with a digital camera. Images are acquired at the maximum resolution of the camera with no zoom. More details are described in the manual for the software.