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Updated: Mar 9, 2026
Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
Published on: December 3, 2019
Isolation of Microglia and Immune Infiltrates from Mouse and Primate Central Nervous System
Thais F Galatro1, Ilia D Vainchtein2, Nieske Brouwer2
1Department of Neurology, School of Medicine, University of São Paulo, São Paulo, 01246903, Brazil.
Abstract:
Microglia are the innate immune cells of the central nervous system (CNS) and play an important role in the maintenance of tissue homeostasis, providing neural support and neuroprotection. Microglia constantly survey their environment and quickly respond to homeostatic perturbations. Microglia are increasingly implicated in neuropathological and neurodegenerative conditions, such as Alzheimer's disease, Parkinson's disease, and glioma progression. Here, we describe a detailed isolation protocol for microglia and immune infiltrates, optimized for large amounts of post mortem tissue from human and rhesus macaque, as well as smaller tissue amounts from mouse brain and spinal cord, that yield a highly purified microglia population (up to 98 % purity). This acute isolation protocol is based on mechanical dissociation and a two-step density gradient purification, followed by fluorescence-activated cell sorting (FACS) to obtain pure microglia and immune infiltrate populations.
Insights
This study presents a new protocol to isolate pure microglia and immune cells from brain tissue. This method is effective for human, macaque, and mouse samples, aiding research into neurodegenerative diseases.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key innate immune cells in the central nervous system (CNS).
- They maintain tissue homeostasis and provide neuroprotection.
- Microglia are implicated in neurodegenerative diseases like Alzheimer's and Parkinson's disease.
Purpose of the Study:
- To develop a detailed protocol for isolating microglia and immune infiltrates.
- To optimize the protocol for various tissue types and species (human, macaque, mouse).
- To achieve high purity of isolated microglia populations.
Main Methods:
- Acute isolation protocol using mechanical dissociation.
- Two-step density gradient purification.
- Fluorescence-activated cell sorting (FACS) for purification.
Main Results:
- The protocol yields highly purified microglia populations (up to 98% purity).
- Optimized for both large post-mortem human/macaque tissue and smaller mouse brain/spinal cord samples.
- Successfully isolates microglia and other immune infiltrates.
Conclusions:
- This protocol provides a reliable method for obtaining pure microglia.
- Facilitates research on microglia's role in CNS homeostasis and disease.
- Applicable across different species and tissue quantities.
