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J Vis Exp. 2010 Jul 6;(41). pii: 2081. doi: 10.3791/2081.

DiOLISTIC labeling of neurons from rodent and non-human primate brain slices.

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1
Section on Neuronal Structure, Laboratory for Integrative Neuroscience, NIH - National Institute of Health, Bethesda, MD, USA.

Abstract

DiOLISTIC staining uses the gene gun to introduce fluorescent dyes, such as DiI, into neurons of brain slices (Gan et al., 2009; O'Brien and Lummis, 2007; Gan et al., 2000). Here we provide a detailed description of each step required together with exemplary images of good and bad outcomes that will help when setting up the technique. In our experience, a few steps proved critical for the successful application of DiOLISTICS. These considerations include the quality of the DiI-coated bullets, the extent of fixative exposure, and the concentration of detergent used in the incubation solutions. Tips and solutions for common problems are provided. This is a versatile labeling technique that can be applied to multiple animal species at a wide range of ages. Unlike other fluorescent labeling techniques that are limited to preparations from young animals or restricted to mice because they rely on the expression of a fluorescent transgene, DiOLISTIC labeling can be applied to animals of all ages, species and genotypes and it can be used in combination with immunostaining to identify a specific subpopulation of cells. Here we demonstrate the use of DiOLISTICS to label neurons in brain slices from adult mice and adult non-human primates with the purpose of quantifying dendrite branching and dendritic spine morphology.

PMID:
20644510
PMCID:
PMC3156079
DOI:
10.3791/2081
[Indexed for MEDLINE]
Free PMC Article
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