Ballistic labeling of pyramidal neurons in brain slices and in primary cell culture

Hailong Li, Kristen A. McLaurin, Charles F. Mactutus, Rosemarie M. Booze

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of pyramidal neurons and dendritic spines, a ballistic labeling technique can be utilized. In the present protocol, pyramidal neurons are labeled with DilC18(3) dye and analyzed using neuronal reconstruction software to assess neuronal morphology and dendritic spines. To investigate neuronal structure, dendritic branching analysis and Sholl analysis are performed, allowing researchers to draw inferences about dendritic branching complexity and neuronal arbor complexity, respectively. The evaluation of dendritic spines is conducted using an automatic assisted classification algorithm integral to the reconstruction software, which classifies spines into four categories (i.e., thin, mushroom, stubby, filopodia). Furthermore, an additional three parameters (i.e., length, head diameter, and volume) are also chosen to assess alterations in dendritic spine morphology. To validate the potential of wide application of the ballistic labeling technique, pyramidal neurons from in vitro cell culture were successfully labeled. Overall, the ballistic labeling method is unique and useful for visualizing neurons in different brain regions in rats, which in combination with sophisticated reconstruction software, allows researchers to elucidate the possible mechanisms underlying neurocognitive dysfunction.

Original languageEnglish
Article numbere60989
JournalJournal of Visualized Experiments
Issue number158
StatePublished - Apr 2020

Bibliographical note

Funding Information:
This work was funded by NIH grants HD043680, MH106392, DA013137, and NS100624.

Publisher Copyright:
© 2020 Journal of Visualized Experiments.


  • Ballistic labeling
  • Confocal imaging
  • Dendritic spine
  • Hippocampus
  • Issue 158
  • Neuroscience
  • Pyramidal neuron
  • Rat

ASJC Scopus subject areas

  • Neuroscience (all)
  • Chemical Engineering (all)
  • Biochemistry, Genetics and Molecular Biology (all)
  • Immunology and Microbiology (all)


Dive into the research topics of 'Ballistic labeling of pyramidal neurons in brain slices and in primary cell culture'. Together they form a unique fingerprint.

Cite this