Quantitative determination of neuronal size and density using flow cytometry

Author(s)
Farrow, L F
Andronicos, N M
McDonald, P G
Hamlin, A S
Publication Date
2021-03-15
Abstract
<i>Background:</i> Recent anthropomorphic disturbances are occurring at an increasing rate leading to organisms facing a variety of challenges. This change is testing the information processing capacity (IPC) of all animals. Brain function is widely accepted to be influenced by a variety of factors, including relative size, number of neurons and neuronal densities. Therefore, in order to understand what drives an animals IPC, a methodological approach to analyze these factors must be established.<br/> <i>New method:</i> Here we created a protocol that allowed for high-throughput, non-biased quantification of neuronal density and size across six regions of the brain. We used the Isotropic Fractionator method in combination with flow cytometry to identify neuronal and non-neuronal cells in the brains of adult rats.<br/> <i>Comparison with existing methods:</i> The results obtained were comparable to those identified using stereological counting methods.<br/> <i>Results:</i> By employing this new method, the number of nuclei in a specific brain region can be compared between replicate animals within an experiment. By calibrating the forward scatter channel of the flow cytometer with size standard beads, neuronal and non-neuronal nuclear sizes can be estimated simultaneously with nuclei enumeration. These techniques for nuclear counting and size estimation are technically and biologically reproducible.<br/> <i>Conclusion:</i> Use of flow cytometry provides a methodological approach that allows for consistency in research, so that information on brain morphology, and subsequent function, will become comparable across taxa.
Citation
Journal of Neuroscience Methods, v.352, p. 1-7
ISSN
1872-678X
0165-0270
Pubmed ID
33484745
Link
Publisher
Elsevier BV
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International
Title
Quantitative determination of neuronal size and density using flow cytometry
Type of document
Journal Article
Entity Type
Publication

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