Potato cyst nematodes (PCN) are notorious pathogens in all major potato production areas worldwide. Mainly due to the low mobility of this soil pathogen, PCN infestations are mostly observed as patches ('foci') that only cover a fraction of the acreage. In-field pre-symptomatic localization of this pathogen is valuable as it would allow for the localized application of control measures. Although the mapping of foci is technically feasible, it is unpractical as it would take the analysis of numerous soil samples. We investigated whether chlorophyll fluorescence (Chl-F) could be suitable as a rapid, non-destructive method for early PCN detection. To this end, the impact of four Globodera pallida densities on the Chl-F of tomato was investigated in a phenotyping greenhouse for 26 days. Furthermore, classical plant performance indicators biomass and root surface area were compared with Chl-F. Thermal dissipation ('NPQ_Lss') and actual photosynthetic rate ('QY_Lss') responded at 1 DPI, while QY_Lss was most sensitive to low PCN infection levels. Chl-F parameters responded more readily to PCN infection than biomass and root surface area. The efficiency of photosystem II (QY_max) and the potential activity of photosystem II (Fv/Fo) initially increased at low PCN infection levels, whereas a sharp decrease was observed at higher infestation levels. Hence, our data suggest that low PCN levels promoted plant performance before becoming detrimental at higher levels. While Chl-F allowed for early and sensitive PCN detection, it remains to be investigated whether these signals can be distinguished from those produced by other below-ground stressors in the field.