“…The two-plate analgesiameter for freely moving mice was used in this study ( Figure 2 ). This prototype device is our own solution, which has several advantages as compared to classical analgesiameters, such as: (1) measuring only thermally-evoked pain, (2) controlling current temperature on each plate of the device using advanced proportional-integral-derivative (PID) algorithms implemented in PLC S7-1200 controller (Siemens, Germany) [ 32 ], which allows the mouse to choose the preferred thermal zone, without being forced by the researcher, (3) rapid and precise regulation of the temperature set-point on each of the two plates of this device enables to establish separate “thermal zones” randomly in each testing session, which significantly limits potential false positive results due to animals’ ability to learn choosing the preferred thermal zone of the device (i.e., a preferred plate) based on external visual cues, (4) the possibility of setting narrow-range thermal zones on each plate, which increases sensitivity of the method; at the same time this allows to measure the effect of a drug on hyperalgesia and allodynia, (5) the measurements are collected from unrestrained animals and therefore this method is less stressful for animals as compared to some standard thermal pain tests (e.g., cold water test and cold plate test), thus offering more reliable data due to a reduction of stress-related behavioral reflex responses of experimental animals (e.g., stress-induced analgesia) [ 33 ] and (6) higher objectivity of measurements (i.e., observer-independent assessment) because time spent by the animal in a given thermal zone and trajectories are recorded by the video camera (GoPro Hero7 Black, San Mateo, CA, USA) and subsequently they are analyzed with the use of mathematical methods of image analysis (deep learning and machine learning) [ 34 , 35 ]. Of note, the camera used for the detection of animals’ movements enables one to assess the effect of test drugs on locomotor activity of mice and this allows one to exclude potential false positive results in the pain test that are due to decreased locomotor activity [ 31 ].…”