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For monitoring the cellular pH in plants, engineered green fluorescent protein (GFP) has been used to indicate cellular pH. Pt-GFP is a pH-reporter protein which has been used ratiometrically in the double excitation mode, according to the fluorescence properties described for Pt-GFP (Schulte et al., 2006), and depending on our previous experiments, we present here a little and efficient protocol for monitoring the cytosolic pH value. In this protocol, root hair cellular pH was monitored in the PtGFP reporter lines. Cellular pH can be obtained according to the calibration curve which was performed in situ by using a series pH buffer.