Can I use the nucleator on saggital sections?

Question

I would like to use the nucleator to measure the volume of the cells that I count in an optical fractionator. From reviewing the stereology literature I see that the nucleator can be used on isotropic or vertical sections. What about sagittal sections? Any help would be appreciated.

Discussion

The nucleator is a technique that can be used to estimate paticle volume. As with all stereological techniques there is a requirement that the probe and the objects interact with some degree of randomness. The word probe refers to the geometric shape interacting with the objects of interest. In the case of the nucleator the probe is a straight line. This straight line represents a random line in the 3D world. Since the tissue is normally sectioned, the possible choices for the 3D line are restricted. Within the section the orientation of the line is restricted to the angles from 0 to 180 degrees.

So how can all of the possible lines that might have run through the original tissue be represented? The answer is that the sections must be cut with random orientation.

A sagittal section is a preferred orientation. Using these sections it is not possible to run any 3D line through the particles being measured. This isn’t a problem if the particles appear the same here as they do in all other section orientation.

This doesn’t mean that the particles have to be spheres. Banana shaped particles work too as long as they are randomly oriented. The problem is that it is difficult to tell if the particles are randomly oriented.

The strict answer to the question is that sagittal sections cannot used. On the other hand, sagittal sections can be used if it is proved that the particles are randomly oriented. Verifying the random orientation of the particles is much more difficult than estimating their volume. The most demanding issue is that this check must be done in all experiments because what is true in the control tissue may not be true in the experimental tissue.

If random selection is broken at any point in the process, “all bets are off.”

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