Fig. 1

a, b Normalised signal of a logarithmic dilution of fixed iRFP expressing A431 cells (a) or iRFP antibody (680RD) (b), suspended in low-melt agarose, and a representative example of each dilution series as seen by each platform. Gating was performed to be able to see the lowest possible dilution above background. A representative example of one of the triplicates imaged in each platform is shown below the graph. Arrows represent the subjective last visible dilution using gating optimised to see the lowest possible signal distinguishable from background. LPT = Li-Cor Pearl Trilogy, BIX = Bruker In-Vivo Xtreme, XVI = Xenogen VivoVision IVIS 200. Quantification was performed in each platform’s own software and results of technical triplicates with standard deviation error bars are shown. Points without error bars have an error too low to be rendered in Prism. c Schematic representation of calliper measurements (blue triangles) and imaging measurements (green triangles) in days after subcutaneous injection of iRFP expressing H1299 cells. d Tumour volume over time as measured by callipers. Error bars represent ± SD, (asterisk indicates Tukey’s multiple comparisons test p < 0.05, NS p > 0.05). e–g Measurements of fluorescent intensity of tumours over time as measured by the XVI (fluorescent intensity: radiant efficiency) (e) BIX (fluorescent intensity: counts) (f) or the LPT (fluorescent intensity: signal) (g). Error bars represent ± SD, (asterisk indicate statistically significant differences, ANOVA, Tukey’s multiple comparisons test p < 0.05, NS indicates no significant difference). h–j Images of the tumour of mouse 2 on each measurement day for the XVI (h), BIX (i) and LPT (j). Images show fluorescent intensity as shown by each platform’s software (false colours mode) overlaid on brightfield images, to show example of tumour growth. Scale bars for each software are presented. XVI was gated to day 17, BIX and LPT were gated to day 14