How to count cells: the advantages and disadvantages of the isotropic fractionator compared with stereology

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• 作者：Suzana Herculano-Houzel (1) (2)
  Christopher S. von Bartheld (3)
  Daniel J. Miller (4)
  Jon H. Kaas (4)
  
  1. Instituto de Ci锚ncias Biom茅dicas ; Universidade Federal do Rio de Janeiro ; Rio de Janeiro ; Brazil
  2. Instituto Nacional de Neuroci锚ncia Translacional ; Minist茅rio de Ci锚ncia e Tecnologia ; Sao Paulo ; Brazil
  3. Department of Physiology and Cell Biology ; University of Nevada School of Medicine ; Reno ; NV 89557 ; USA
  4. Department of Psychology ; Vanderbilt University ; Nashville ; TN 37240 ; USA
  
• 关键词：Numbers of neurons ; Brain size ; Isotropic fractionator ; Stereology ; NeuN
• 刊名：Cell and Tissue Research
• 出版年：2015
• 出版时间：April 2015
• 年：2015
• 卷：360
• 期：1
• 页码：29-42
• 全文大小：4,886 KB
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Proteomics
  Molecular Medicine
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0878

文摘

The number of cells comprising biological structures represents fundamental information in basic anatomy, development, aging, drug tests, pathology and genetic manipulations. Obtaining unbiased estimates of cell numbers, however, was until recently possible only through stereological techniques, which require specific training, equipment, histological processing and appropriate sampling strategies applied to structures with a homogeneous distribution of cell bodies. An alternative, the isotropic fractionator (IF), became available in 2005 as a fast and inexpensive method that requires little training, no specific software and only a few materials before it can be used to quantify total numbers of neuronal and non-neuronal cells in a whole organ such as the brain or any dissectible regions thereof. This method entails transforming a highly anisotropic tissue into a homogeneous suspension of free-floating nuclei that can then be counted under the microscope or by flow cytometry and identified morphologically and immunocytochemically as neuronal or non-neuronal. We compare the advantages and disadvantages of each method and provide researchers with guidelines for choosing the best method for their particular needs. IF is as accurate as unbiased stereology and faster than stereological techniques, as it requires no elaborate histological processing or sampling paradigms, providing reliable estimates in a few days rather than many weeks. Tissue shrinkage is also not an issue, since the estimates provided are independent of tissue volume. The main disadvantage of IF, however, is that it necessarily destroys the tissue analyzed and thus provides no spatial information on the cellular composition of biological regions of interest.