In Vivo Bioluminescence Imaging of Gene Delivery

Mice injected with the PFV-luciferase vector were subsequently imaged 13 and 49 days after injection by whole-body bioluminescence imaging (IVIS) (Caliper Life Sciences, Hopkinton, MA, USA) as described previously.⁵⁵ (link) Those that received PFV-EGFP were sacrificed on day 11 and examined for direct EGFP expression using a stereoscopic fluorescence microscope (MZ16F, Leica Microsystems, Wetzlar, Germany) as described previously.⁵⁶ (link) Images were captured using a digital microscope camera (DFC420, Leica Microsystems, Milton Keynes, UK) and software (Image Analysis, Leica Microsystems). Mice that received intracerebral injections of double-stranded DNA (dsDNA) plasmid were imaged 5 days and 11 days post-injection, with luciferase activity normalized to the bioluminescent signal produced by replicate 1 of the uninjected group.

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Counsell J.R., Karda R., Diaz J.A., Carey L., Wiktorowicz T., Buckley S.M., Ameri S., Ng J., Baruteau J., Almeida F., de Silva R., Simone R., Lugarà E., Lignani G., Lindemann D., Rethwilm A., Rahim A.A., Waddington S.N, & Howe S.J. (2018). Foamy Virus Vectors Transduce Visceral Organs and Hippocampal Structures following In Vivo Delivery to Neonatal Mice. Molecular Therapy. Nucleic Acids, 12, 626-634.

Publication 2018

MZ16F DFC420

Camera Dsdna Fluorescence microscope Luciferase Mice Microscope Plasmid Replicate Vector

Corresponding Organization : South African Medical Research Council

Other organizations : Great Ormond Street Hospital, University of Würzburg, National Hospital for Neurology and Neurosurgery, TU Dresden

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Variable analysis

independent variables

Injection of PFV-luciferase vector
Injection of PFV-EGFP
Injection of double-stranded DNA (dsDNA) plasmid

dependent variables

Bioluminescence imaging (IVIS) at 13 and 49 days after injection
Direct EGFP expression using a stereoscopic fluorescence microscope on day 11
Luciferase activity 5 days and 11 days post-injection

control variables

Bioluminescent signal produced by replicate 1 of the uninjected group

controls

Uninjected group as a negative control for the luciferase activity measurement

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