Ultrastructural morphometry using dual axes tangential scale: a technical revelation.

ABSTRACT

While performing ultrastructural morphometry, under or over estimation of ultrastructural size could be avoided by using accurate measuring devices. Biological investigators have always relied on conventional linear scale for the baseline measurement of ultrastructural size parameters on electron micrographs to project the dimensions of intracellular organelles or tissue components. Since it was not possible to measure decimal fractions of mm with linear scale, a 'dual axes tangential scale' has been designed for measuring ultrastructural image parameters on electron micrographs with an accuracy of 0.1 mm to minimize the error in finally computed size of ultrastructural component. In an exercise using 'dual axes tangential scale' and 'conventional linear scale', measurement of glomerular basement membrane thickness (GBMT) as orthogonal intercepts across the GBM revealed a 'coefficient of variation' at 4.4% with dual axes tangential scale as compared to 'coefficient of variation' at 10.9% with linear scale, expressing superiority of dual axes tangential scale over linear scale. Use of mathematical formula rather than nomogram has been preferred. However, 'slide guide, ultrastructure size calculator' could also be used for discerning ultrastructural size after measurement with dual axes tangential scale.