D luciferin bioluminescence substrate

The procedure has been described elsewhere (Genovese et al., 2017 (link)). A 4.7T Bruker Biospec (BrukerBioSpin), equipped with 35 mm innerdiameter volume coil and 6 cm inner-diameter gradients, was used for imaging the animals. A fast acquisition with relaxation enhancement (RARE) sequence with TR/TE of 2,000/38 ms, matrix size 256× 192, 0.75 mm slice thickness, 0.25 mm slice gap, 4× 3 cm FOV, 101 kHz bandwidth, 3 NEX was used for acquired in coronal and axial geometries a multi-slice T2-weighted images. To reduce the respiratory motion the axial scan sequences were gated. IVIS-100 imaging system was used for the detection of luciferase activity. Mice were injected with d-luciferin bioluminescence substrate (Perkin Elmer), according to manufacturer’s instructions, and imaged 5 min after the injection. In ex vivo experiments mice were sacrificed and tissues harvested 5 min after luciferin injection. The living Image 4.3 software (Perkin Elmer) was used for analysis of the images after acquisition.
For ex vivo fluorescence imaging animals were sacrificed and imaged using a Leica MZ125 Stereo Zoom microscope and a DFC450C camera.

Animals were imaged on a 4.7T Bruker Biospec (Bruker BioSpin) equipped with 6-cm inner-diameter gradients and a 35-mm inner-diameter volume coil. Multi-slice T2-weighted images were acquired in coronal and axial geometries using a rapid acquisition with relaxation enhancement (RARE) sequence with TR/TE of 2,000/38 ms, matrix size 256 × 192, 0.75-mm slice thickness, 0.25-mm slice gap, 4 × 3-cm FOV, 101-kHz bandwidth, 3 NEX. Axial scan sequences were gated to reduce respiratory motion. Detection of luciferase activity was performed in an IVIS-100 imaging system. Five minutes before the procedure, mice were injected intraperitoneally with d-luciferin, bioluminescence substrate (Perkin Elmer) according to the manufacturer’s instructions. Living Image 4.3 software (Perkin Elmer) was used for analysis of the images after acquisition.