Figure 1.
Macro flow chart for the “Intravital_Microscopy_Toolbox” program.
Starting from the file selection, the image dissimilarity score is computed for each frame in the video. Main processing options are identified with grey shading.
Figure 2.
Calculation of dissimilarity scores using the automatic reference frame generation (RFG) option, and subsequent removal of artifactual frames from the video sequence.
(a) Graph plotting of the total number of frames in a 1000-frame video against the dissimilarity score, identified as ‘square difference’ score on the y-axis. (b) Relative frequency distribution of the number of frames per dissimilarity score. (c) Graph showing the frame-by-frame analysis of a 1000-frame video sequence. Artifactual frames were eliminated (identified by red crosses) using a cutoff percentile of 60% (defined by the horizontal blue line). Major artifacts (shown as black dots) are typically artifacts resulting in a full field distortion of the image, while minor artifacts (shown as green dots) are small glitches in frames that do not impair the image interpretation. (d) Images corresponding to local maxima (upper panels) and minima (lower panels) in the first 200 frames of the video sequence. Note that frames corresponding to a local maximum are associated with heavy artifacts, while frames corresponding to a local minimum are typically very similar to the reference frame. All images are derived from Video S3.
Figure 3.
Comparison of x–y transformations efficiency to correct for motion artifacts induced by peristaltism in the murine myenteric plexus.
(a) Artifactual displacement from frame to frame for an object either located in the center (object identified by the white cross in Video S5) or periphery (white circle in Video S5) of the field of view. The ‘Sum’ indicates the total artifactual displacement of an object in the video sequence. (b) Plots illustrating the relative motion of selected objects within the field of view. ***p<0.001, **p<0.01, and *p<0.05, one-way ANOVA with Bonferroni post-test.