Microglia Activation Assay Service

Microglia, the central nervous system's resident immune guardians, hold the key to understanding neuroinflammation, neurodegenerative diseases like Alzheimer's disease (AD), Parkinson's disease (PD), and Multiple Sclerosis (MS), and the intricate processes of brain injury repair. Our in vitro microglia activation assay service offers researchers an efficient and precise method for dissecting the complexities of microglial activation. For further information regarding the products and services provided, project-specific consultation, and pricing, please submit an inquiry here.

Introduction

Microglial activation is not a simple on/off switch, but rather a complex process involving a multitude of signaling pathways and molecular mechanisms. These pathways orchestrate the release of cytokines, chemokines, and other signaling molecules that influence the inflammatory response and the fate of surrounding cells. Understanding the intricate interplay of these signaling cascades is essential for deciphering the precise role of microglia in both neuroprotection and neurodegeneration.

Fig.1 Microglia activation receptors and pathways.¹

What We Provide?

• Isolation and Purification of Microglia

Our system leverages cutting-edge techniques to ensure exceptional microglial purity. We employ advanced methods like magnetic activated cell sorting (MACS) and flow cytometry (FACS). This multi-layered approach guarantees a microglial population exceeding 95% purity, minimizing the confounding effects of contaminating cell types and maximizing the reliability of your data.

Maintaining microglia in a state that closely resembles their in vivo environment is crucial for accurate and translatable research. Our system utilizes a chemically defined culture medium, carefully formulated with essential factors like TGF-β2 and M-CSF. This approach avoids the non-specific activation often associated with traditional serum-based media, ensuring that your microglia remain in a near-homeostatic state.

Fig.2 Isolation of CD11b⁺ cells.²

• Building Robust Microglial Activation Models

LPS model: Standardized concentrations induce proinflammatory phenotypes as evidenced by morphology (amoebic changes) and CD86 labeling.

Aβ model: Aβ peptide treatment simulates AD pathology, detects reactive oxygen species (ROS) and superoxide release, and correlates with neurotoxicity assessment.

Cytokine induction: IFNγ combined with LPS synergistic activation simulates the neuroinflammatory cascade response and quantifies the intensity of inflammation through indicators such as TNF-α, IL-1β and iNOS.

• Microglial Activation Status Analysis

Morphological Analysis: Quantitative assessment of microglial morphology, including cell size, process length and branching, and changes in cell shape, provides valuable information about their activation state and functional polarization. We utilize advanced imaging techniques and software for precise morphometric analysis.

Cytokine and Chemokine Profiling: Measurement of secreted cytokines and chemokines provides a direct assessment of microglial activity and their influence on the surrounding microenvironment. We employ multiplex assays and ELISA to quantify a broad panel of inflammatory mediators.

Gene Expression Analysis: Analysis of gene expression profiles reveals the molecular pathways and transcriptional changes associated with microglial activation. We utilize RNA sequencing and qPCR to identify differentially expressed genes and characterize the functional state of microglia.

Protein Expression Analysis: Assessment of protein levels provides insights into the translational regulation of microglial activation. We employ Western blotting and flow cytometry to quantify key proteins involved in signaling pathways and effector functions.

Functional Assays: Direct measurement of microglial functions, such as phagocytosis, migration, and ROS production, provides a comprehensive assessment of their activation status and potential impact on neuronal health.

Our services seamlessly integrate the entire research process, from cell isolation to functional analysis. We combine literature-verified methodologies with customized solutions to provide reliable data support for neuroimmunology research. If you require further technical details or collaboration, please do not hesitate to contact us.

References

1. Qin, Jun, et al. "Microglia activation in central nervous system disorders: A review of recent mechanistic investigations and development efforts." Frontiers in Neurology 14 (2023): 1103416. Distributed under Open Access license CC BY 4.0, without modification.
2. Nikodemova, Maria, and Jyoti J. Watters. "Efficient isolation of live microglia with preserved phenotypes from adult mouse brain." Journal of neuroinflammation 9 (2012): 1-10. Distributed under Open Access license CC BY 2.0, without modification.