However, as a result of this sectioning process, we lose one dimension, and therefore we lose important qualitative and quantitative information. Conventional light and transmission electron microscopy (TEM) rely on this principle. To get insight into the fine structure of three-dimensional biological objects, the classical approach is to use microscopes to investigate nearly two-dimensional sections produced by microtomes from small samples taken from these objects. This pertinacious problem may be termed the “dimension trap”. In the context of microscopy, there is a related problem regarding the misinterpretation of what we see. And although we are aware of this problem, they still work. Many optical illusions are based on our misinterpretation of two-dimensional figures as representations of three-dimensional objects. The “dimension trap” in microscopy and its solution by stereology The symbiosis of stereology and 3D imaging thus provides the unique opportunity for unbiased and comprehensive quantitative characterization of the three-dimensional architecture of the lung from macro to nano scale. Here, we review those stereological principles that are of particular relevance for 3D imaging and provide an overview of applications of 3D imaging-based stereology to the lung in health and disease. micro-computed tomography, confocal microscopy and volume electron microscopy. Stereology is, therefore, perfectly suited to be combined with 3D imaging techniques which cover a wide range of complementary sample sizes and resolutions, e.g. Thus, three-dimensionality is a built-in property of stereological sampling and measurement tools. It takes into account the fact that, in traditional microscopy such as conventional light and transmission electron microscopy, although one has to rely on measurements on nearly two-dimensional sections from fixed and embedded tissue samples, the quantitative data obtained by these measurements should characterize the real three-dimensional properties of the biological objects and not just their “flatland” appearance on the sections. Stereology is the method of choice for the quantitative assessment of biological objects in microscopy.
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