Fully-stratified organotypic cultures of the REK cell line were pretreated with MTX or with media alone for 72 hr at the concentration indicated, followed by incubation with ALA (1 mM, 4 hr) prior to frozen-sectioning. (A) Frozen sections visualized for PpIX by confocal microscopy. Note the increased intensity of the bright PpIX-specific signal in cultures pretreated with methotrexate. (B) Quantification of PpIX signal. Integrated fluorescence was analyzed from 6 confocal images per condition, using IPLab image-processing software. (*), significant difference from control, p < 0.005 level.