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Related Experiment Video

Updated: Aug 28, 2025

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

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

Published on: December 3, 2019

Using mechanical homogenization to isolate microglia from mouse brain tissue to preserve transcriptomic integrity.

Shawn Herron¹, Jean-Christophe Delpech², Charlotte Madore²

¹Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA, USA.

|September 15, 2022

Summary

This study introduces a new method for isolating microglia, a type of brain cell, using mechanical homogenization instead of heat and enzymes. This preserves the cells' natural gene expression for more accurate research.

Keywords:

Cell isolation Cell separation/fractionation Classification Description: Cell biology Flow cytometry/Mass cytometry Gene expression Molecular biology Neuroscience RNAseq

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Last Updated: Aug 28, 2025

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

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Published on: December 3, 2019

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Author Spotlight: Enhancements in Gene Expression Regulation Research

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Published on: September 15, 2023

Area of Science:

Neuroscience
Immunology
Cell Biology

Background:

Microglia isolation is crucial for studying brain health and disease.
Current enzymatic methods at 37°C can alter microglia gene expression, affecting research outcomes.
Accurate microglia transcriptomics are essential for understanding neurological conditions.

Purpose of the Study:

To develop an optimized protocol for isolating microglia with minimal transcriptomic changes.
To provide a reliable method for obtaining high-quality microglia for gene expression analysis.
To overcome the limitations of enzymatic dissociation in microglia research.

Main Methods:

Mechanical homogenization of mouse brain tissue using Dounce homogenizer.
Single-cell suspension preparation.
Microglia isolation via Percoll gradient and flow cytometry.

Main Results:

The optimized protocol successfully isolates microglia.
Isolated microglia show preserved gene expression patterns, avoiding heat-induced artifacts.
Mechanical homogenization prevents transcriptomic alterations common in enzymatic methods.

Conclusions:

This mechanical homogenization protocol offers a superior alternative for microglia isolation.
The method ensures the integrity of microglia gene expression for reliable downstream assays.
This technique is vital for advancing research in neuroinflammation and brain disorders.