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Color Movies with VoxBlast: Flood Fill of Microtubules Near a Mitochondrial Mass
This movie illustrates several features in VoxBlast and shows the power of the flood fill option. This dataset was captured with a BioRad confocal microscope (MRC 600) using a 100x, 1.4 N.A. objective. The data was then deconvolved with MicroTome from VayTek to remove residual haze and sharpen the microtubules. Then, the data set was rendered in VoxBlast.
The research question related to the arrangement of the microtubules around the mitochondrial mass. It is known that the mitochondrial mass does not have a membrane. The question was, then, are the microtubules outside the mitochondrial mass contiguous or discontiguous with the microtubules inside the mitochondrial mass?
To address this issue, a single microtubule outside the mass was chosen. In VoxBlast, a single seed was place in that microtubule. The flood fill feature was used to flood fill that microtubule. The flood fill process jumped to another microtubule where it touched the first. Then, a third microtubule was filled. This process continued until all the microtubules that were contiguous with the first seed were filled. VoxBlast changed the color of the filled microtubules and reported the mass of all filled microtubules.
Finally, a movie loop was made of the flood filled dataset. The movie loop reveals that the microtubules around the mitochondrial mass were filled. However, the microtubules inside the mass were not filled. This arrangement is obvious in the 3D movie loop. It would have been impossible to trace all the connections in the 2D image series by hand. The flood fill was able to answer the question quickly. The results were considered preliminary and further research is needed to confirm them. The data is courtesy of David Gard, University of Utah.
Click here to download a larger version of this movie loop.
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