Argon plasma jet (Ar-PJ) has been widely used in clinical medicine; however, the cellular effects of Ar-PJ therapy applying to living tissues have not been clarified yet. It is necessary to investigate cellular responses to Ar-PJ in establishing guidelines on the therapeutic use of Ar-PJ. Interestingly, in the Ar-PJ-treated cells, the fragmented mitochondria, a typical cellular stress indicator, were discovered even in the cells located in the live zones (1∼3 zones). Using microscopic measurements of the mitochondrial length, we found that the fragmented mitochondria were mainly in the zones 1 and 2, the closest to the direct exposure point of Ar-PJ. Whereas, the mitochondria in the zone 4 retained their lengths to normal. This quantitative measurement of mitochondrial morphology was combined with the color scores of the polymerizable supramolecular (PS) sensor in diagnostic categories. The results demonstrate that the mitochondrial length (0.98∼3.94 μm) is inversely proportional to the PS sensor color scores (87∼0) in the zones 1∼4. On the combination of these three diagnostic parameters, the effective range of Ar-PJ for cellular responses was determined: the zones 1∼3, the color scores 87∼12 and the mitochondrial lengths 0.98∼2.57 μm. Our study is the first demonstration of mitochondrial fragmentation in response to Ar-PJ and the first attempt to establish the diagnostic guideline for Ar-PJ therapies by combinations with biological, physical and chemical aspects. Thus, this study will make great advances in the field of bioplasma applications.