3D Animated Reconstructions
Curiosity is a wonderful thing. The two images to the left are recontructions of Sensitive Fern spores (Onoclea sensibilis L.). There was no immunolabeling or staining of any kind performed. Plant cell walls autofluoresce, among other naturally occurring, usually structural, biological materials. A z-series was collected with a BioRad Radiance 2000 confocal on a Nikon Eclipse 200 inverted light microscope. The z-series was then loaded into VoxBlast, and the two movies on the left were created. Notice all the fine detail that was collected. The apparent "hole" on one side of the structure is an artifact due to the inability of the signal from the bottom of the object to penetrate through the upper layers of material.
These two movies were made using the same technique as above. In this case, we're looking at a Louse claw, preserved in a typical histology slide, that you might find in a high school biology class. Here, the autofluorescent material is the chitin found in the insects exoskeleton.
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This movie shows a macrophage engulfing a bead. From the initial image, as you would see through the microscope, the interaction between the bead and the cell is not clear. However, by taking a z-series, and rotating the three dimensional volume, you can clearly see the interaction. The movie itself took about 10 minutes to generate and illustrates the usefulness of 3-D imaging.
Stills from 3D reconstructions
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The stills shown above were created from the same data set as the louse claw movies. These two images have a different appearance because they were created in BitPlane's more powerful software, Imaris and Surpass. This software is not only capable of handling larger data sets, but it also allows for fly-throughs of the data as seen in the image on the right. In both of these stills, green represents the cuticle, while the red is residual muscle that wasn't removed during the potassium hydroxide treatment used to clear insect bodies for microscopy. The image on the left appears to show a connection point where the muscle anchors to the cuticle. (where the red and bright green points come together just to the right of the center of the image.)