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Date: | Mon, 24 Oct 1994 14:54:30 -0400 |
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This is how I figured out a rough method for data collection with a
BioRad MRC 600 for volume rendering with VoxelView.
We are using Nikon 60X NA 1.4 objective, 1 1/2 coverslips, and glycerol /
PBS/ n-propyl gallate mounting medium.
I expected the Z axis to be incorrect because of index of refraction
problems, but this is not what I found.
A user brought cell cultures to our facility for confocal. One of the
stains was for f-actin. There were a lot of dividing cells. Viewing from
above, conventionally or w/ single optical section, cells in
metaphase appeared to be circular regardless of orientation and uniform in
size. Thus, we assumed the cells were close to spherical. We set the Z
step to a multiple of the X,Y resolution and interpolated accordingly
(e.g. X, Y 0.1 um, Z step 0.3 um, interpolate twice). With 25 um diameter
cells, this takes a long time!
-Michael C.
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