Before starting XF analysis, cells were inoculated into the wells of XF cell culture microplates. A bicarbonate-free medium was used in the analysis. Up to four drugs can be added to the dosing hole of the analysis probe plate for automatic dosing during analysis. First, the analysis probe board is placed in the analyzer to automatically calibrate the optical sensor. Then, insert the cell culture plate into the instrument.

Generally, the baseline (basic) OCR and ECAR rates are measured three or four times before the inhibitor, agonist, substrate or compound is added through the feed port. OCR is reported in pmol/min and ECAR is reported in mpH/min. Then, the compounds preloaded in the dosing port of the probe plate are sequentially pneumatically added to the culture medium of each well. After slow mixing, OCR and ECAR were measured several times.

XF measurement in hippocampus is a non-destructive measurement method, so the metabolic rate of the same cell group can be measured repeatedly when four different drugs are added to each well in turn.

The total analysis time is generally 60 to 90 minutes. After XF analysis is completed, other types of biological analysis, such as cell activity, can be carried out on the same plate. In order to maintain the normal physiological function of cells, a temperature control system is used to maintain the cell environment at 37℃ (or other preset temperature).