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Open Access Highly Accessed Open Badges Correspondence

The problem of pseudoreplication in neuroscientific studies: is it affecting your analysis?

Stanley E Lazic

Author Affiliations

Cambridge Computational Biology Institute, Department of Applied Mathematics and Theoretical Physics, University of Cambridge, Cambridge, CB3 0WA, UK

Oxford Centre for Collaborative Applied Mathematics, Mathematical Institute, University of Oxford, Oxford, OX1 3LB, UK

Current address: F. Hoffmann-La Roche AG, In Silico Sciences - Statistics, 4070 Basel, Switzerland

BMC Neuroscience 2010, 11:5  doi:10.1186/1471-2202-11-5

Published: 14 January 2010



Pseudoreplication occurs when observations are not statistically independent, but treated as if they are. This can occur when there are multiple observations on the same subjects, when samples are nested or hierarchically organised, or when measurements are correlated in time or space. Analysis of such data without taking these dependencies into account can lead to meaningless results, and examples can easily be found in the neuroscience literature.


A single issue of Nature Neuroscience provided a number of examples and is used as a case study to highlight how pseudoreplication arises in neuroscientific studies, why the analyses in these papers are incorrect, and appropriate analytical methods are provided. 12% of papers had pseudoreplication and a further 36% were suspected of having pseudoreplication, but it was not possible to determine for certain because insufficient information was provided.


Pseudoreplication can undermine the conclusions of a statistical analysis, and it would be easier to detect if the sample size, degrees of freedom, the test statistic, and precise p-values are reported. This information should be a requirement for all publications.