The SPR signal in living cells reflects changes other than the area of adhesion and the formation of cell constructions

Surface plasmon resonance (SPR) sensors detected large angle of resonance (AR) changes, when RBL-2H3 rat mast cells were cultured and activated on a sensor chip. Here, we demonstrated that PAM212 mouse keratinocytes also showed a large change in AR, when EGF-stimulated. We explored these changes due to intracellular reactions, through the relationship between the AR and the area of cell adhesion, using confocal microscopy for RBL-2H3 cells and PAM212 cells. The effect of Mycalolide B and Toxin B, inhibitors for cell motility, on AR was observed using RBL-2H3 cells. Measuring AR in the presence of various numbers of non-stimulated cells demonstrated that AR and cell density were proportional. However, the AR increase in response to antigen was 35% higher than that expected by solely an increase of the cell adhesion area. Moreover, the AR with PAM212 cells decreased following a transient increase in response to EGF, whilst the area of cell adhesion remained at an increased level. Furthermore, the treatment of RBL-2H3 cells with either Mycalolide B or Toxin B slightly inhibited, but never abolished the AR increase induced by antigen. These treatments abolished all morphological changes, including ruffling and the increase of cell adhesion area observed by light microscopy. These results suggest that AR changes reflect intracellular events rather than changes in the size of the area to which cells adhere.