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VayTek, Inc.
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Embedding Traces in Sectioned Slices
of a Chick Embryo

The ultimate goal of this project was to view the course of an artery in a chick embryo. There were several steps in this process which are outlined in the next few pages. First the embryo had to be grown, prepared, sliced and mounted. Next, the slices had to be imaged and stored to disk. After the images were enhanced and aligned, traces were made around the artery in each of the slices. Finally, the slices were embedded in the original image and viewed as three dimensional rotations in VoxBlast. Each of these steps is illustrated in more detail on the following pages.

A chick embryo was prepared and sliced with a microtome. The individual slices (about 60) were mounted on a single glass slide. The slide was mounted on an Olympus BHT microscope. A 40x, .75 N.A. lens was used to view the slices. A Dage CCD 72 camera captured individual images of the slices. Each image was manually position under the microscope. A stack of 60 images was collected and stored to disk.

Since the images were noisy and contained background contamination, several image enhancement procedures were applied. First, a background image was subtracted from every image in the stack. Second, each image in the stack was deconvolved using VayTek's proprietary deconvolution program HazeBuster. Finally, gamma and contrast enhancement were used to stretch the histogram and create the optimal image. In the images above the top left image shows a raw image. The top right image shows the results of deconvolution and background subtraction.

Because the images were not registered, an alignment procedure had to be performed. The Align module in VoxBlast was used for this purpose. Each image was examined and aligned manually using landmarks and a reference image. After the manual operation was complete, the module performed an automated alignment to fine tune the process. The bottom left image shows a side view before alignment; the bottom right image shows a side view after alignment. You can see a better indication of the alignment procedure by looking at the next page which has movie loops illustrating the effects of the alignment.

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