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Related Experiment Video

Updated: May 10, 2026

Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
09:49

Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing

Published on: December 3, 2019

Microglia isolation from adult mouse brain.

Jae-Kyung Lee1, Malú G Tansey

  • 1Emory University School of Medicine, Atlanta, GA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 2, 2013
PubMed
Summary

This study presents a new protocol for isolating intact microglia from adult mouse brains. The method ensures high yields for various downstream applications, including functional assays and gene expression analysis.

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Area of Science:

  • Neuroscience
  • Immunology
  • Cell Biology

Background:

  • Microglia isolation from adult mouse brain is challenging, often yielding low cell counts.
  • Existing protocols are insufficient for obtaining high-quality microglia for comprehensive analysis.

Purpose of the Study:

  • To develop and validate a robust protocol for isolating intact microglia from adult mouse brains.
  • To ensure isolated microglia are suitable for diverse downstream applications.

Main Methods:

  • Enzymatic dissociation using dispase II, papain, and DNase I.
  • Mechanical dissociation followed by Percoll gradient centrifugation for cell separation.
  • Optimization of dissociation and separation steps for adult mouse brain tissue.

Main Results:

  • Successful isolation of intact microglia from adult mouse brains with improved yields.
  • Demonstrated suitability of isolated microglia for flow cytometry, RNA isolation, and functional assays.
  • Validated microglia integrity and viability for downstream molecular and functional analyses.

Conclusions:

  • The described protocol provides a reliable method for obtaining high-quality adult mouse microglia.
  • This advancement facilitates further research into microglia function and roles in the adult brain.
  • The protocol supports a wide range of applications, from gene expression studies to functional assays.

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Last Updated: May 10, 2026

Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
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Published on: September 6, 2024