Living Image 3.1

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11. 3D Reconstruction of Sources 11.2 3D Reconstruction of Fluorescent Sources Set Up & Acquire an Image Sequence on the IVIS ® Spectrum Generate the Surface Topography Set the FLIT Parameters & Reconstruct the Sources 142 Use the imaging wizard to setup the image sequence required for FLIT analysis. (For more details on the imaging wizard, see page 23.) If you plan to manually set up the sequence, Table 11.2 shows the recommended image sequence. Acquire the images using transillumination on the IVIS Spectrum using the same excitation and emission filters from at least four source locations. Table 11.2 Example image sequence for FLIT analysis Image Type Source Location First location Subsequent locations Photographic ✓ Use previous photo Structured light ✓ Fluorescent ✓ ✓ For instructions, see page 137. After the surface is generated, the 3D reconstruction of the light sources can proceed. 1. In the tool palette, click FLIT 3D Reconstruction. Analyze tab Source 01 corresponds to the first image of the sequence, source 02 corresponds to the second image, and so on 2. In the Properties tab, make a selection from the Tissue Properties drop-down list. “Muscle” is usually the best choice of tissue properties for general in vivo reconstructions. Note: The internal medium index of refraction is automatically entered when you select a tissue.
Living Image ® Software User’s Manual Parameters tab Properties tab 3. In the Params tab, confirm the parameter defaults or enter new values. For more details on the parameters, see Appendix H, page 238. It is recommended that you start with the default parameter settings. If necessary, fine tune the settings in the parameters tab. For more details on the parameters and options, see Appendix H, page 233. 4. In the Analyze tab, select the source locations to include in the analysis and click Reconstruct. - Figure 11.1, page 141 shows example 3D reconstruction results 143
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Living Image 3.1

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