Living Image 3.1

Living Image ® Software User’s Manual
12
Troubleshooting Guide
12.1 DLIT/FLIT Analysis Troubleshooting
Table 12.1 DLIT/FLIT analysis
Issue Solution
Large voxels (> 2 mm) The Ndata>Nvoxels is required for an overdetermined problem. It is possible that there are not
enough data (N Surface). This can occur because it is desired that all wavelengths (DLIT) and all
trans-illumination source positions (FLIT) are represented in the data equally. In the DLIT case,
if the signal in a short wavelength is very low, then the sampled data can be too few for sufficient
refinement of the voxel gridding.
– To increase the data to be included, determine the [photons/sec/cm2/sec] level for ~30 counts
on the low signal image (this is the level of noise) using Image Adjust in the Tool Palette. Enter
in the [photons/sec/cm2/sec] value into the 'Minimum Radiance' column next to the
wavelength/image number in the 'Analyze Tab' in the DLIT/FLIT 3D Reconstruction menu of
the Tool Palette.
It is best to include the low signal data as it adds constraining information to the solution.
Note: Using the Imaging Wizard should reduce the occurrence of low signal images in the data
sequences.
No sources in solution In DLIT or FLIT, this can occur if the surface is not correct. That is, if a surface is imported into
the 3D View from another source other than from the Structured Light Analysis.
In FLIT, if you choose an emission Source Spectrum in the Properties Tab (an Advanced User
feature) which is incorrect for the fluorophore of interest, this can also result in no sources.
In FLIT, if there is very little signal detected at the surface and the 'Background' box is checked
in the Params tab, then it is possible that only the background fluorescence is simulated.
Surfaces are spiky The most common source of spiky surfaces are folds in the animal skin or animal fur, which
corrupt the desired smooth lines projected on the animal from the laser galvanometer.
– Adjusted the 'Path Averaging Size' in the 'Surface Topography' menu in the Tool Palette can
help reconstruct an improved surface
– Smoothing the surface by using the 'Smooth' feature in the tool palette Surface Topography
menu can also help improve the surface.
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