In vivo imaging was performed with an IVIS Lumina Spectrum, which allows for the analysis of bioluminescence and near-infrared signals, and was analyzed with LivingImage software (PerkinElmer). Four 8, 11, and 15 days after EAE immunization, 150 μl of XenoLight RediJect Inflammation Probe (40 mg/ml; PerkinElmer), which recognizes myeloperoxidase activity, was injected intraperitoneally. Bioluminescence was captured 5, 10, and 15 min after injection in 9–10 animals per genotype to assess individual differences in XenoLight kinetics. During imaging, mice were kept under 1–1.5% isoflurane anesthesia. The IVIS settings were as follows: Epi-bioluminescence, emission filter open, excitation filter block, fstop = 1, binning = 8, focus B = 6.5 cm, exposure = 60 sec. The regions of interest (ROI) i.e., the injection sites, were identified by using the automatic detection tool in LivingImage. For each mouse, the two time points with the maximum total counts of bioluminescence in the ROIs were used to statistically analyze the genotype differences.
Lumina spectrum
Manufactured by PerkinElmer
The Lumina Spectrum is a high-performance luminescence spectrometer designed for accurate and reliable optical measurements. It features a compact and modular design, allowing for flexible configuration to meet diverse laboratory requirements.
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2 protocols using lumina spectrum
1
Bioluminescence Imaging of Myeloperoxidase Activity in EAE
2
Paw Inflammation Imaging in Mice
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In vivo imaging of paw inflammation was done with an IVIS Lumina Spectrum as described34 and bioluminescence signals were analysed with Living Image software (Perkin Elmer). Fourteen days after induction of paw inflammation, 150 μL XenoLight RediJect Inflammation Probe (40 mg/mL; PerkinElmer) that recognizes myeloperoxidase activity was injected intraperitoneally and bioluminescence of the hind paw was captured 5, 10 and 15 minutes after injection in eight animals per genotype. During the imaging procedure, mice were kept under 1%‐1.5% isoflurane anaesthesia. The IVIS settings were as follows: Epi‐bioluminescence, emission filter open, excitation filter block, fstop 1, binning 8, focus B 6.5 cm, exposure 60 seconds. Regions of interest (ROI) that is the ipsilateral paws were set to software‐aided automatic detection. The contralateral paw served as control. For each mouse, the two maximum time points of the total counts of bioluminescence in ROIs were used for statistical analysis of genotype differences.
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