Biomedical Engineering Reference
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tissue ribbon is line-sampled at 300-nm resolution, near the Nyquist rate for an
ideal optical resolution of (0.77)(532 nm) = (0.80 NA) = 512 nm. Based on this, the
total data size (assuming one byte per voxel) comes out to 20 TB (at half the reso-
lution in each dimension, it would be
2.5 TB). The tissue ribbon can be sampled
at 11 mm/s by line sampling at 44 kHz (180 Mbps), the camera maximum (Dalsa
CT-F3-4096 pixels). Sampling the 2.9-km tissue ribbon requires 265,000s = 73 hr.
Because mice brains are not cubical, stage return takes time, and soon we add 50%
overhead, resulting in
∼
100 hr.
Figures 2.4 and 2.5 show typical data that can be obtained using the KESM [9].
Nissl staining dyes the RNA in the cytoplasm of all neurons and the DNA in cell
bodies in all cells. However, the dendritic arbors and axons remain unstained.
∼
Figure 2.5
Golgi data from KESM. The stack of images generated by KESM can be viewed from the
side of the stack (resectioning). A single resectioned plane is shown at the top. Golgi stain results in
sparse data, so oftentimes it is easier to see familiar structures by overlaying multiple planes (middle).
A single neuron can be observed when approximately 300 planes are overlayed (bottom). (Adapted
from [9].)
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