Abstract
Design-based stereology is a quantification method to obtain a precise and unbiased estimate of the total number of cells (or any other objects) in a well-defined region of interest. There are two comparable stereological counting methods, (a) the Optical Fractionator and (b) the Nv:Vref method. Due to the adherence to strict stereological protocol, the Optical Fractionator is the most unbiased and preferable stereological method. However, the Nv:Vref method can be an alternative when tissue availability is limited. Both methods use systematic random sampling (SRS) techniques to account for the inhomogeneous nature of biological tissue. Here we describe the criteria for a successful and accurate stereological study, using human brain tissue.
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Notes
- 1.
Although this chapter focuses on design-based stereology in the context of human brain research, the same principles apply to small and large animal studies.
- 2.
Systematic random sampling (SRS) is regular sampling (using a known interval) that begins with a randomized start. SRS is the basis for design-based stereological sampling methods, such as the Optical Fractionator .
- 3.
The volume associated with the counting space or box.
- 4.
- 5.
The coefficient of error (CE) is the precision of a population size estimate. It is calculated as the standard deviation of the sample divided by the mean of the sample. This is used as a measurement of the accuracy of estimated total cell count .
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Mehrabi, N.F., Singh-Bains, M.K., Waldvogel, H.J., Faull, R.L.M. (2018). Stereological Methods to Quantify Cell Loss in the Huntington’s Disease Human Brain. In: Precious, S., Rosser, A., Dunnett, S. (eds) Huntington’s Disease. Methods in Molecular Biology, vol 1780. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7825-0_1
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