324. Development of Bioluminescence Tomography for Three-Dimensional Real Time Imaging of Gene Expression
Year: 2006
Type: article
Abstract: Introduction. Non-invasive monitoring of gene expression in living animals is an active area of molecular imaging and has proven useful to identify real time gene transfer. Improvements in optical imaging techniques have lead to the mounting luciferase reporter systems. Standard 2D bioluminescence limits the interpretation of gene expression in vivo. Bioluminescence tomography (BLT), the 3D extension of bioluminescence imaging, offers a new tomographic method for molecular imaging. BLT captures the spatial distribution of luciferase light signal in three dimensions, which provides a powerful tool for studies such as cancer imaging, cell trafficking and protein function. Methods. 22-gram, male BALB-C mice were used for in vivo experiments. Mice tail veins were injected with 1 ml and 2 ml of 10ug/ml luciferase DNA expression plasmid in PBS. Twenty-four hours after injections, animals were anesthetized and administered D-luciferin at 140mg/kg i.p.. Bioluminescence images were acquired 5 min after D-luciferin administration. The mice were immersed in an imaging chamber filled with 0.5% intralipid as a coupling medium. The stage was rotated 3X at 90° and the emitted bioluminescence was collected for 3 minutes using a cooled CCD camera. Results. Figs. 1a and 1b (20ug and 10ug dose respectfully) show the imaging chamber containing the mouse under measurement in the absence and presence of intralipid solution. Fig. 1c shows the bioluminescence from a single 90o data point. Figs. 2b and d (20ug and 10ug dose respectfully) were obtained from a single cross section of the mouse at the z-axis=20mm. Figs. 2a and c (20ug and 10ug dose respectfully) are reconstructed 3D images at multiple planes showing the complete bioluminescence distribution. Conclusion. The purpose of these studies were to validate the algorithm for converting bioluminescence into a three dimensional images and the instrumentation used to obtain the images, hence the justification for the two DNA doses. The results provide more detail with regard to real time gene expression in that the lower plasmid DNA dose appears in the lower lobe of the liver whereas the higher DNA dose yields more expression in the upper lobes of the liver. We are investigating the use of this technology for imaging brain tumors using RG2 cells permanently transformed to constituitively express luciferase. The anticipation is that this can be used to assay for in vivo gene transfer to brain tumors. View Large Image Figure ViewerDownload Hi-res image Download (PPT)
Source: Molecular Therapy
Institution: University of Florida
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W2030038817