Ten female CD1 nu/nu mice (6-8 weeks old, 25-30g) were subcutaneously injected with 1×106 cells from a human colorectal carcinoma cell line, SW1222, on their right flank. These cells were previously engineered in-house to express the luciferase gene for bioluminescence imaging (BLI) 76. Mice were housed for 8 days to allow tumours to develop. Prior to 5-ALA administration, each tumour was imaged using BLI and 20 Raman spectra were collected via direct contact measurement with the skin at numerous locations (n = 18-20) from both the tumour and the control flank for each mouse using the theranostic system described using the 785 nm laser with a 100 mW power output and 1 second integration time. Five mice received a 50 mg/kg tail vein injection of a 15 mg/mL 5-ALA solution while five control mice received no injection. At 1, 2, and 4 hours post 5-ALA administration, tumours were imaged using BLI and Raman spectra were obtained from the tumour and the control flanks of all mice as described. After the final Raman spectral acquisitions, mice were treated with the 633 nm PDT laser with a fluence rate of 50 mW/cm2 and a total fluence of 30 J/cm2. Mice were rehoused and monitored for 6 days post PDT. At 1, 3, and 6 days post PDT, tumours of control and PDT mice were reimaged using BLI, and Raman spectra were obtained from the tumour and the control flanks of all mice as described. At 6 days post PDT, the 5 PDT mice were re-injected with 50 mg/kg of a 15 mg/mL 5-ALA solution to enable quantification of PPIX tumoural uptake, but no additional PDT was performed. All mice were subsequently sacrificed by cervical dislocation, in accordance with local regulations, and tumours excised for PPIX quantification.

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