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

Journal of Visualized Experiments : JoVE
Gail K SeaboldVeronica A Alvarez

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...Continue Reading

References

Apr 16, 2003·Methods : a Companion to Methods in Enzymology·Jaime GrutzendlerWen-Biao Gan
Feb 24, 2006·Proceedings of the National Academy of Sciences of the United States of America·Ko-Woon LeePaul Greengard
Oct 20, 2007·Trends in Biotechnology·John A O'Brien, Sarah C R Lummis
Mar 6, 2009·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Hao-wei ShenPeter W Kalivas
Aug 29, 2009·Journal of Neuroscience Methods·M Diana NeelyAriel Y Deutch
Feb 12, 2010·Cold Spring Harbor Protocols·Wen-Biao GanJeff W Lichtman

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Citations

Jul 29, 2011·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Verginia C Cuzon CarlsonKathleen A Grant
Sep 3, 2013·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Mark V WilcoxVeronica A Alvarez
Feb 4, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Alice DobiVeronica A Alvarez
May 28, 2013·Cell and Tissue Banking·April T DavenportJames B Daunais
Jul 20, 2014·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Robert F RoscoeRosemarie M Booze
May 23, 2014·Frontiers in Neuroanatomy·Connie ChengLaurie C Doering
Apr 27, 2016·Neuroscience·Scott A WongAaron J Gruber
Apr 12, 2020·Proceedings of the National Academy of Sciences of the United States of America·Tomoya NakayamaTakashi Yoshimura

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Methods Mentioned

BETA
dissecting
confocal microscopy
transgenic

Software Mentioned

DiOLISTICS

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