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On Measuring Section Thickness

Published online by Cambridge University Press:  14 March 2018

Ron Doole*
Affiliation:
University of Oxford

Extract

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Section thickness can be measured by placing beads of some kind on the top and bottom surfaces of the section. This is then a simple parallax problem.

Imagine the specimen in cross section. If there are two particles, one vertically above the other they are separated by the film thickness T. Tilt the film through an angle A and in plan view the particles will separate by a distance D. This can also be extended to account for two particles not vertically above each other but I'll stick to the easy case for the explanation.

Take two negatives one at zero tilt and one at tilt of A and measure the separation D. The thickness can be calculated by T=D/sinA.

The direction of the tilt axis must be known for the measurements and it is easy to see that the larger the tiit angle and the more accurately the separation is measured, the more accurate the measurement will be. Tilt at both positive and negative angles to get a more accurate result.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2001

References

Hawes, C.R., 1981. Application of high voltage electron microscopy to botanical ultrastructure. Micron, Vol. 12 No. 3. pp 227257.Google Scholar
Nankivell, J.F. 1963. The theory of electron stereo microscopy. Optik, Vol.20, pp 171198.Google Scholar