DiOLISTIC labeling of neurons from rodent and non-human primate brain slices.
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.
Seabold
Gail K
GK
Section on Neuronal Structure, Laboratory for Integrative Neuroscience, NIH - National Institute of Health, Bethesda, MD, USA.
Daunais
James B
JB
Rau
Andrew
A
Grant
Kathleen A
KA
Alvarez
Veronica A
VA
D000818:Animals; D019470:Biolistics / Q000379:methods*; D001921:Brain / Q000166:cytology*; D001923:Brain Chemistry; D003712:Dendrites / Q000737:chemistry; D000882:Haplorhini; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D009474:Neurons / Q000737:chemistry / Q000166:cytology*; D014414:Tungsten / Q000737:chemistry
D014414:Tungsten