Home › Immunology › Cell-based Assay to Study Antibody-mediated Tau Clearance by Microglia
Immunology JoVE (Open Access) Citable · DOI

Cell-based Assay to Study Antibody-mediated Tau Clearance by Microglia

DOI: 10.3791/58576-v
What you'll learn
  • ✓ Perform a cell-based tau uptake assay using BV-2 microglia cells
  • ✓ Apply FACS to quantify antibody-mediated tau clearance mechanisms
  • ✓ Analyze flow cytometry data to assess anti-tau antibody efficacy
Protocol

Here we describe a cell-based assay to quantitatively assess tau uptake by microglia with the aim of creating an investigational tool to better characterize the mechanisms of action of anti-tau antibodies.

Difficulty
intermediate
Total time
~4–6 hours (cell culture incubation and FACS acquisition per batch)
Model organism
BV-2 cells (immortalized mouse microglia)
Biosafety
BSL-1

Steps

1
Set up tau uptake assay with FACS readout

Prepare BV-2 microglia cells and establish conditions for tau protein uptake in the presence or absence of anti-tau antibodies. Configure cell culture and incubation parameters to enable quantitative measurement by flow cytometry.

▶ 00:37
2
Perform FACS analysis on treated samples

Acquire and gate flow cytometry data from tau-treated microglia, measuring fluorescent tau uptake in single cells. Apply compensation and gating strategies to isolate relevant cell populations.

▶ 04:33
3
Quantify antibody-mediated tau uptake results

Compare tau uptake levels between control and antibody-treated BV-2 cells using FACS data. Calculate fold-changes or mean fluorescence intensity to determine the efficacy of anti-tau antibodies in promoting microglial tau clearance.

▶ 05:41
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