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Updated: Aug 15, 2025
Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
Protocol for assessing phagocytosis activity in cultured primary murine microglia
Elsie Layman1, Jennifer Michelle Parrott2, Hye Young Lee1
1Department of Cellular and Integrative Physiology, University of Texas, Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Abstract:
In this protocol, we describe steps to assess inflammation-induced cell response in cultured primary murine microglia through the analysis of fluorescent bead phagocytosis. We detail primary murine mixed glial cell culture preparation followed by microglia-specific isolation. Further, we describe treatment with lipopolysaccharide (LPS) to induce phagocytosis of fluorescent beads, followed by quantitative analysis using fluorescent imaging and Fiji - ImageJ software. For complete details on the use and execution of this protocol, please refer to Parrott et al.1.
Insights
This protocol details how to measure inflammation in mouse microglia cells by tracking their uptake of fluorescent beads after stimulation with lipopolysaccharide (LPS). This method quantifies cellular responses to inflammatory stimuli.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the brain, crucial for understanding neuroinflammation.
- Assessing microglia's inflammatory response is vital for studying neurological disorders.
Purpose of the Study:
- To provide a detailed protocol for quantifying inflammation-induced phagocytosis in primary microglia.
- To establish a reproducible method for analyzing microglia cell response.
Main Methods:
- Primary murine mixed glial cell culture and microglia isolation.
- Stimulation with lipopolysaccharide (LPS) to induce inflammatory response.
- Phagocytosis assay using fluorescent beads and quantitative analysis via fluorescent imaging (Fiji/ImageJ).
Main Results:
- The protocol enables quantitative assessment of microglia phagocytic activity.
- Lipopolysaccharide (LPS) effectively induces an inflammatory response measurable by bead uptake.
Conclusions:
- This protocol offers a reliable method for studying microglia inflammatory responses in vitro.
- The described technique is valuable for neuroinflammation research and drug screening.
