DiOLISTIC Labeling of Neurons from Rodent and Non-human Primate Brain Slices

Section on Neuronal Structure, Laboratory for Integrative Neuroscience, NIH - National Institute of Health, Bethesda, MD, USA.
Journal of Visualized Experiments (Impact Factor: 1.33). 07/2010; 6(41). DOI: 10.3791/2081
Source: PubMed


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.

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    • "ontal cortex and hippocampus ( Adalbert and Coleman , 2013 ) . Here , we showed confocal images of the DiI - labeled dendrite segments from the brain slices of both the control groups and silibinin - treated groups . Two important characteristics to look for are a sparse labeling pattern and the ability to identify individual cellular components ( Seabold et al . , 2010 ) . The cell body was medium - sized , and the dendrites were densely covered with spines . The pictures demonstrated the degree of the increase in the average number of dendritic spines on a single neuron from the mice in the silibinin - treated groups . Our results showed that the average density of dendritic spines on each neuron , w"
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    • "Typical applications of this technique include the study of neuronal morphology during development and altered development in neurological disorders (Bruce et al., 1997; Braun and Segal, 2000; Smith et al., 2009; Li et al., 2010). This dye can be applied to a variety of cell types, live or fixed tissue (Terasaki et al., 1994), as well as diverse species such as rodents, primates, and zebrafish (Gan et al., 2000; O’Brien and Lummis, 2006; Seabold et al., 2010; Arsenault and O’Brien, 2013). In slice preparations, DiI labeling is commonly known as “DiOlistic labeling,” in which beads coated with the lipophilic dye are “ballistically” ejected with a gene gun on to brain tissue (Lo et al., 1994). "
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