Design-based stereology: Planning, volumentry and sampling are crucial steps for a successful study. Systems biology in 3D space - enter the morphome. often difficult to identify in mice using standard dissection techniques. This method requires specialist equipment, including a microscope with a motorized stage driven. using standard stereological methods such as volume fractions of tissue. Cutting a fine figure - on the use of thin sections in electron microscopy to quantify autophagy. Based on these principles, stereology is considered to be the gold-standard for cell counting, as with proper sampling parameters, the results derived from this technique are an unbiased and accurate estimate of primordial follicle numbers 4, 35. number of objects requires a 3-D (volume) probe and therefore the disector technique. Any new development in microscopy broadens the potential applications of stereology. The term stereology is used in estimating quantities in the. As such, it is independent from particular imaging techniques but universally applicable.
The number of points multiplied by the area associated with one of those points on the grid lattice will be an estimate of profile area of that organelle.įor 3D quantitation we analyse consecutive serial sections to obtain data about organelle volumes and number. Stereology is deeply rooted in stochastic geometry and can be used as a toolbox for sampling and measuring biological objects in microscopy. A regular array of equally spaced points is used to count points that land over the areas of interest. An example would be the estimation of the area of an organelle by point counting. Depending on the desired parameter these probes can be points, lines or planes. In a second step we apply stereological estimators in form of geometrical probes onto the sampled micrographs/locations and count “events” between those probes and the underlying ultrastructural features. In most cases this sampling scheme is carried out by a systematic uniform random placement throughout the specimen which gives every location an equal chance of being included in the sample. In an initial step a rigorous sampling scheme ensures unbiased selection of the areas/locations of interest. We will now use this technique to further define the growth of the ureteric tree in vitro, under both normal culture conditions, and in the presence of various levels of specific. Repeat estimates of total tree length vary by just 1-2. We use well established stereological tools to obtain unbiased estimates of cellular features like sizes, volumes or numbers of organelles and particles. Preliminary data shows the total length estimates obtained with the technique to be highly reproducible.