<i>In vitro</i> and <i>ex vivo</i> detection of control_IgG_680, A1_Mab_680 and MMP_750 fluorescence by IVIS imaging in EAE.

(A) In vitro assessment of control IgG_680 shows a signal at the appropriate wavelength (row 1). In experiments that used both MMP_750 and A1_Mab_680 fluorescence in the same animal, there was no overlap in signal (rows 2 and 3). (B) In vivo imaging: four groups of mice (non-EAE, pre-disease, peak disease and chronic disease) were injected with either control_IgG_680 (rows 1 and 5) or A1_Mab_680 and MMP_750 (rows 2–4, 6 and 7) 24 hours prior to sacrifice. Clinical scores were obtained on the day of sacrifice. Row 1: Spinal cord control_IgG_680 imaging shows signal in two of five animals at the peak disease stage and marginal signal at other time points. Row 2: Spinal cord A1_Mab_680 imaging shows signal predominantly at the peak and chronic stages of EAE. Row 3: Spinal cord MMP_750 imaging closely parallels the A1_Mab_680 imaging. Row 4: Merged images of rows 3 and 4 show overlap between A1_Mab_680 and MMP_750 and a caudal to rostral shift of both signals. Row 5: Brain control_IgG_680 imaging shows modest signal in the same animals as the spinal cord imaging. Rows 6 and 7: Brain A1_Mab_680 and MMP_750 imaging (respectively) show increased signal in brain.