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More details on appropriate stereological parameters for various animal models of human lung disease are given in.
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Derived from these parameters, there are others like mean particle volume ( \(\overline\)). Modified after Įxpress the alterations in your target compartment as changes in stereological parameters such as volume (V), surface area (S), length (L) or number (N). Note that in this cascade sampling design, the phase of interest at one level (red) becomes the reference phase (red + green) at the next level. The total volume of type II cells, V(typeII), is then obtained as V(typeII) = V(lung). The volume fraction of type II cells within alveolar septum, V V(typeII/alvsep), is estimated by point counting at a medium electron microscopic magnification. The volume fraction of alveolar septum within parenchyma, V V(alvsep/par), is estimated by point counting at a medium light microscopic magnification (compare Fig. At a low light microscopic magnification, the volume fraction of parenchyma within lung, V V(par/lung), is estimated by point counting. Total lung volume, V(lung), is the product of t and the total cut area of the apical side of all slices (shown in red). These slices serve two purposes: They can be used for the estimation of total lung volume by the Cavalieri estimator (Step 3) as well as for sampling of tissue blocks for light and electron microscopy (see Fig. At the macroscopic level, the lung is cut completely into horizontal slices of thickness t, starting at a random position between 0 and t (arrow).
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by systematic uniform random sampling, have to be applied (Step 4). At each level, from macroscopic via light microscopic to electron microscopic, the principles of unbiased sampling, e.g. Illustration of a cascade sampling design in the lung, using the estimation of the total volume of alveolar type II cells in a lung as an example. You just have to be willing to change a few lab habits regarding the processing of lung tissue for “standard histology”. For (almost) every task there is an appropriate tool. The production of data is straightforward.
STEREOLOGY COUNTING RULES HOW TO
The basic principles how to use stereology can be learnt in a few hours. Therefore, you find a little glossary that defines common stereological terms in Table 2. These tools have names that may sound unfamiliar to you. Rest assured that this is actually an advantage because it provides a solid scientific foundation (from stochastic geometry) for what you will be doing. Don’t be deterred by the mathematical theory of stereology (although we have to confess that some of the original articles we reference in this primer are a tough read). It aims to help you to do better next time, in particular by planning ahead for a quantitative assessment of lung micro-structure.
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What went wrong? If you don’t know, this primer is for you. Sadly, without these data the journal is not willing to accept your paper for publication. After a few emails have been exchanged, this expert tells you that your study cannot be rescued at this stage, because all you have available for analysis is a few paraffin sections taken arbitrarily from each mouse lung without knowledge of the volume of the fixed lung, but these alone are not suitable for stereological analysis (compare Table 1 for common problems). You approach one of the leading scientists in the field of lung stereology and ask for help. There is even an official research policy statement by the American Thoracic Society (ATS) and the European Respiratory Society (ERS) about this. What now? After a brief search, you find out that there exists something called stereology (in case the reviewer didn’t already mention it) that is considered the gold standard to quantitate lung architecture in microscopy. One (or more) of the reviewers is asking for a formal quantification of the histological findings in your animal model. After a few weeks, you receive the reviewers’ comments. Your nice and elaborate experimental study demonstrating that your favorite molecule is essential in the pathogenesis of a common lung disease is ready for submission.