In this study, a dual microsensing electrochemical probe for measuring oxygen (O2) and pH levels was developed based on a dual recessed Pt disk electrode (each disk diameter, 10 μm) with the use of two Ag/AgCl reference electrodes (one for each disk of the dual electrode). One of the recessed Pt disks of the dual electrode was electrodeposited with a porous Pt layer and then coated with a hydrophobic photocured polymer (partially fluorinated epoxy diacrylate, abbreviated as FED). The Pt-FED covered disk was used as an amperometric O2 sensor and exhibited a linear current increase that was proportional to the PO2 level (partial O 2 pressure) with high sensitivity (168.4 ± 33.8 pA mmHg -1) and fast response time (t90% = 0.17 ± 0.05 s). The other recessed Pt disk was electrodeposited with an IrO2 layer. The potential between the IrO2 deposited electrode and the Ag/AgCl reference electrode produced a reliable Nernstian response to pH changes (58.3 ± 1.5 mV pH-1) with a t90% of 0.43 ± 0.09 s. The sensor displayed high stability in the in vitro organ tissue measurements for at least 2.5 h. By using the developed dual O2/pH microsensor as a probe tip for scanning electrochemical microscopy, the two-dimensional images of the location-dependent PO2 and pH levels were simultaneously acquired and could be used to assess the surface of a rat kidney tissue slice. When compared to the corresponding medullary levels, both PO2 and pH were observed to be higher in the cortex area, while the modest level gradient was observed near the cortex-medulla border. This finding suggests that there is a direct relationship between the tissue O2 supply/consumption and pH, which is mainly determined by metabolite, such as CO2, production.