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Published on: October 18, 2016
High Content Imaging and Quantification of Microglia Phagocytosis In Vitro
Christopher A McPherson1, Kiran Kelly-Rajan1,2, Christian Lefebvre d'Hellencourt3
1Mechanistic Toxicology Branch, Division of Translational Toxicology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA.
Abstract:
Microglia function as the tissue-specific resident macrophages of the nervous system, performing immune and non-immune functions. These functions are critical to development and to maintain homeostasis in the nervous system throughout the lifespan, and during brain injury or disease. One method by which microglia maintain homeostasis is phagocytosis of aberrant proteins, extracellular debris, synapses, or apoptotic cells. Phagocytic function can be changed by environmental or genetic risk factors that affect microglia. These protocols present a rapid and simple in vitro high-content imaging protocol for studying phagocytosis in the murine microglia BV-2 cell line. High-content imaging and analysis enable versatility of the assay, which can be used to test multiple experimental conditions, or as a screening tool. © 2023 Wiley Periodicals LLC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA. Basic Protocol 1: Phagocytosis of fluorescently labeled particles Basic Protocol 2: Examining modifications to phagocytosis by test substances Basic Protocol 3: High content imaging and analysis of phagocytic cells.
Insights
This study introduces a fast, in vitro high-content imaging method to analyze microglia phagocytosis. This technique allows researchers to efficiently study how various factors impact microglial immune functions in the nervous system.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are the resident immune cells of the central nervous system, essential for neural development, homeostasis, and response to injury or disease.
- A key microglial function is phagocytosis, the process of engulfing cellular debris, pathogens, and other unwanted materials.
- Dysregulation of microglial phagocytosis is implicated in various neurological disorders.
Purpose of the Study:
- To present a rapid and simple in vitro high-content imaging protocol for studying phagocytosis in the murine microglia BV-2 cell line.
- To provide a versatile assay for testing multiple experimental conditions and serving as a screening tool for phagocytic function.
Main Methods:
- Development of an in vitro high-content imaging protocol using the BV-2 murine microglia cell line.
- Protocol 1: Quantifying phagocytosis of fluorescently labeled particles.
- Protocol 2: Assessing modifications to phagocytosis induced by test substances.
- Protocol 3: Detailed high-content imaging and analysis of phagocytic cells.
Main Results:
- The described protocol enables rapid and simple assessment of microglial phagocytic activity.
- High-content imaging allows for versatile analysis, accommodating multiple experimental variables.
- The assay can be effectively utilized as a screening tool for identifying modulators of phagocytosis.
Conclusions:
- This high-content imaging protocol offers an efficient method for studying microglial phagocytosis in vitro.
- The developed assay is valuable for both detailed mechanistic studies and large-scale screening of factors affecting microglial function.
- This approach contributes to a better understanding of microglia's role in maintaining nervous system health and disease.
