On counting and counting errors
Abstract
Counting objects in histological sections is often a necessary, sometimes an unexpected part of a research project. The recent literature shows that the subject of counting is of particular interest to readers of the Journal of Comparative Neurology but that it is also contentious and difficult. Even a brief review of past issues of the Journal shows that there are many misconceptions about counting and that there remain issues that have received little or no attention. Counts are subject to many errors. Some reports include readily recognizable errors, others fail to include all of the information that is needed for an evaluation of their accuracy. This review is above all a plea for adequate information about the methods used for counts in all publications. It serves to help those who are new to quantitative methods in histology; it considers some of the basic issues arising for anyone undertaking counts, or reviewing manuscripts that include counts. In particular, it considers recently introduced or re-introduced counting methods that depend on accurate measures along the axis perpendicular to the plane of the sections, and looks at the difficulties inherent in these measures.
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An Abercrombie correction was applied to all cell counts to correct for overcounting in profile sections.
“An Abercrombie (1946) correction was applied to all cell counts in order to correct for overcounting in profile sections (see Guillery, 2002).”
From Quantitative mapping reveals age and sex differences in vasopressin, but not oxytocin, immunoreactivity in the rat social behavior neural network (2017) · cites this paper as Guillery, 2002
Abercrombie corrections based on the size of cell nuclei in each subject were performed to estimate numbers of hypocretin and MCH neurons.
“To estimate accurately the numbers of hypocretin and MCH neurons in each section, we performed Abercrombie corrections based on the size of the cell nuclei in each subject.13”
From Loss of hypocretin (orexin) neurons with traumatic brain injury (2009) · cites this paper as [13]
Cites
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von Bartheld, C. S., Bahney, J., & Herculano‐Houzel, S. (2016) · Journal of Comparative Neurology · doi:10.1002/cne.24040 · Cited by 1,063 · see citing works
Xu, X., Roby, K. D., & Callaway, E. M. (2010) · Journal of Comparative Neurology · doi:10.1002/cne.22229 · Cited by 429 · see citing works
Characterization of Kiss1 neurons using transgenic mouse models
Cravo, R. M., Margatho, L. O., Osborne-Lawrence, S., Donato, J., Atkin, S., Bookout, A. L., Rovinsky, S., Frazão, R., Lee, C. E., Gautron, L., et al. (2011) · Neuroscience · doi:10.1016/j.neuroscience.2010.11.022 · Cited by 353 · see citing works
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Mouse cortical inhibitory neuron type that coexpresses somatostatin and calretinin
Xu, X., Roby, K. D., & Callaway, E. M. (2006) · The Journal of Comparative Neurology · doi:10.1002/cne.21101 · Cited by 185 · see citing works
Loss of hypocretin (orexin) neurons with traumatic brain injury
Baumann, C. R., Bassetti, C. L., Valko, P. O., Haybaeck, J., Keller, M., Clark, E., Stocker, R., Tolnay, M., & Scammell, T. E. (2009) · Annals of Neurology · doi:10.1002/ana.21836 · Cited by 185 · see citing works
“To estimate accurately the numbers of hypocretin and MCH neurons in each section, we performed Abercrombie corrections based on the size of the cell nuclei in each subject.(13) We then calculated the total numbers of hypocretin and MCH neurons by multiplying the cell counts per section by the section sampling frequenc…”
MethodParallel preoptic pathways for thermoregulation
Yoshida, K., Li, X., Cano, G., Lazarus, M., & Saper, C. B. (2009) · The Journal of Neuroscience · doi:10.1523/jneurosci.2643-09.2009 · Cited by 160 · see citing works
A hypothalamic circuit for the circadian control of aggression
Todd, W. D., Fenselau, H., Wang, J. L., Zhang, R., Machado, N. L., Venner, A., Broadhurst, R. Y., Kaur, S., Lynagh, T., Olson, D. P., et al. (2018) · Nature Neuroscience · doi:10.1038/s41593-018-0126-0 · Cited by 154 · see citing works
Distribution of progesterone receptor immunoreactivity in the fetal and neonatal rat forebrain
Quadros, P. S., Pfau, J. L., & Wagner, C. K. (2007) · Journal of Comparative Neurology · doi:10.1002/cne.21427 · Cited by 112 · see citing works
Segregation of feedforward and feedback projections in mouse visual cortex
Berezovskii, V. K., Nassi, J. J., & Born, R. T. (2011) · Journal of Comparative Neurology · doi:10.1002/cne.22675 · Cited by 84 · see citing works
Loss of Gsx1 and Gsx2 function rescues distinct phenotypes in Dlx1/2 mutants
Wang, B., Long, J. E., Flandin, P., Pla, R., Waclaw, R. R., Campbell, K., & Rubenstein, J. L. R. (2013) · Journal of Comparative Neurology · doi:10.1002/cne.23242 · Cited by 85 · see citing works
DiBenedictis, B. T., Nussbaum, E. R., Cheung, H. K., & Veenema, A. H. (2017) · Journal of Comparative Neurology · doi:10.1002/cne.24216 · Cited by 74 · see citing works
“An Abercrombie (1946) correction was applied to all cell counts in order to correct for overcounting in profile sections (see Guillery, 2002).”
MethodChalfin, B. P., Cheung, D. T., Muniz, J. A. P. C., de Lima Silveira, L. C., & Finlay, B. L. (2007) · Journal of Comparative Neurology · doi:10.1002/cne.21406 · Cited by 61 · see citing works
Somatostatin, tau, and β-amyloid within the anterior olfactory nucleus in Alzheimer disease
Saiz-Sanchez, D., Ubeda-Bañon, I., de la Rosa-Prieto, C., Argandoña-Palacios, L., Garcia-Muñozguren, S., Insausti, R., & Martinez-Marcos, A. (2010) · Experimental Neurology · doi:10.1016/j.expneurol.2009.06.010 · Cited by 63 · see citing works
Metabolic Effects of Chronic Sleep Restriction in Rats
Vetrivelan, R., Fuller, P. M., Yokota, S., Lu, J., & Saper, C. B. (2012) · Sleep · doi:10.5665/sleep.2200 · Cited by 61 · see citing works
Bingham, B., & Viau, V. (2008) · Endocrinology · doi:10.1210/en.2007-1796 · Cited by 44 · see citing works
Disney, A. A., & Reynolds, J. H. (2014) · Journal of Comparative Neurology · doi:10.1002/cne.23456 · Cited by 40 · see citing works
“Brightness and contrast settings were chosen by using a live color image.”
MethodGenetic tracing of subpopulation neurons in the prethalamus of mice (<i>Mus musculus</i>)
Delaunay, D., Heydon, K., Miguez, A., Schwab, M., Nave, K., Thomas, J. L., Spassky, N., Martinez, S., & Zalc, B. (2009) · Journal of Comparative Neurology · doi:10.1002/cne.21904 · Cited by 33 · see citing works
Disney, A. A., Alasady, H. A., & Reynolds, J. H. (2014) · Brain and Behavior · doi:10.1002/brb3.225 · Cited by 28 · see citing works
- Authors
- Guillery, R. W.
- Affiliations
- University of Wisconsin–Madison (US)
- Corresponding author
- Guillery, R.W.
- Venue
- Journal of Comparative Neurology
- Publisher
- Wiley
- Published
- 2002-04-08
- Volume
- 447
- Issue
- 1
- Pages
- 1-7
- ISSN
- 0021-9967 (ISSN-L)
- 1096-9861
- OpenAlex
- W1968759033
- Cited by
- 343
- References
- 2
- Citations by year
- 2025: 3 · 2024: 5 · 2023: 4 · 2022: 5 · 2021: 5 · 2020: 7 · 2019: 3 · 2018: 10 · 2017: 12 · 2016: 10 · 2015: 19 · 2014: 14 · 2013: 26 · 2012: 19
- FWCI
- 1.13
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Comparative Neurology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01ey011494-04)
- National Eye Institute (EY11494, R01EY012936, EY 12936)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Health Sciences › Medicine › Anatomy › Medical and Biological Sciences
- Physical Sciences › Mathematics › Applied Mathematics › Point processes and geometric inequalities
- Keywords
- counting errors, quantitative histology, counting methods, measurement accuracy, 3D measurement
- MeSH terms
- Animals, Cell Count, Humans, Microscopy, Microtomy, Neuroanatomy, Research Design, Observer Variation, Selection Bias, Artifacts, Optics and Photonics
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