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Stereological Methods to Quantify Cell Loss in the Huntington’s Disease Human Brain

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Book cover Huntington’s Disease

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1780))

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. 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. 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. 3.

    The volume associated with the counting space or box.

  4. 4.

    An unbiased stereological method that estimates the volume of a structure from individual parallel cross-sectional areas (typically sections) employing Cavalieri’s principle [7] Typically, the areas are estimated using a point-counting method [8].

  5. 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 .

References

  1. Thu DC, Oorschot DE, Tippett LJ et al (2010) Cell loss in the motor and cingulate cortex correlates with symptomatology in Huntington’s disease. Brain 133:1094–1110

    Article  PubMed  Google Scholar 

  2. Kim EH, Thu DC, Tippett LJ et al (2014) Cortical interneuron loss and symptom heterogeneity in Huntington disease. Ann Neurol 75:717–727

    Article  CAS  PubMed  Google Scholar 

  3. Nana AL, Kim EH, Thu DC et al (2014) Widespread heterogeneous neuronal loss across the cerebral cortex in Huntington's disease. J Huntingtons Dis 3:45–64

    PubMed  Google Scholar 

  4. Mehrabi NF, Waldvogel HJ, Tippett LJ et al (2016) Symptom heterogeneity in Huntington’s disease correlates with neuronal degeneration in the cerebral cortex. Neurobiol Dis 96:67–74

    Article  PubMed  Google Scholar 

  5. Singh-Bains MK, Tippett LJ, Hogg VM et al (2016) Globus pallidus degeneration and clinico-pathological features of Huntington’s disease. Ann Neurol 80:185–201

    Article  CAS  PubMed  Google Scholar 

  6. Schmitz C, Hof P (2005) Design-based stereology in neuroscience. Neuroscience 130:813–831

    Article  CAS  PubMed  Google Scholar 

  7. Cavalieri B, Lombardo-Radice L (1966) Geometria degli indivisibili di Bonaventura Cavalieri, vol 5. Unione Tipografico-Editrice Torinese, Torino

    Google Scholar 

  8. Gundersen H, Jensen E (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147:229–263

    Article  CAS  PubMed  Google Scholar 

  9. Waldvogel HJ, Curtis MA, Baer K et al (2006) Immunohistochemical staining of post-mortem adult human brain sections. Nat Protoc 1:2719–2732

    Article  CAS  PubMed  Google Scholar 

  10. West MJ, Gundersen H (1990) Unbiased stereological estimation of the number of neurons in the human hippocampus. J Comp Neurol 296:1–22

    Article  CAS  PubMed  Google Scholar 

  11. Slomianka L, West MJ (2005) Estimators of the precision of stereological estimates: an example based on the CA1 pyramidal cell layer of rats. Neuroscience 136:757–767

    Article  CAS  PubMed  Google Scholar 

  12. West MJ (1999) Stereological methods for estimating the total number of neurons and synapses: issues of precision and bias. Trends Neurosci 22:51–61

    Article  CAS  PubMed  Google Scholar 

  13. Andersen B, Gundersen H (1999) Pronounced loss of cell nuclei and anisotropic deformation of thick sections. J Microsc 196:69–73

    Article  PubMed  Google Scholar 

  14. Gundersen HJG, Jensen EBV, Kieu K, Nielsen J (1999) The efficiency of systematic sampling in stereology—reconsidered. J Microsc 193:199–211

    Article  CAS  PubMed  Google Scholar 

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Author notes

  1. Nasim F. Mehrabi and Malvindar K. Singh-Bains contributed equally with all other contributors.

Authors and Affiliations

  1. Centre for Brain Research, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand

    Nasim F. Mehrabi, Malvindar K. Singh-Bains, Henry J. Waldvogel & Richard L. M. Faull

  2. Department of Anatomy and Medical Imaging, University of Auckland, Auckland, New Zealand

    Nasim F. Mehrabi, Malvindar K. Singh-Bains, Henry J. Waldvogel & Richard L. M. Faull

Authors
  1. Nasim F. Mehrabi
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  2. Malvindar K. Singh-Bains
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  3. Henry J. Waldvogel
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  4. Richard L. M. Faull
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Corresponding author

Correspondence to Richard L. M. Faull .

Editor information

Editors and Affiliations

  1. Brain Repair Group, Cardiff University, Cardiff, United Kingdom

    Sophie V. Precious

  2. Brain Repair Group, Cardiff University, Cardiff, United Kingdom

    Anne E. Rosser

  3. Brain Repair Group, Cardiff University, Cardiff, United Kingdom

    Stephen B. Dunnett

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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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  • DOI: https://doi.org/10.1007/978-1-4939-7825-0_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7824-3

  • Online ISBN: 978-1-4939-7825-0

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