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Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro.

Nicholas D Condon1,2, Jennifer L Stow2, Adam A Wall2

  • 1Institute for Molecular Biosciences (IMB) Microscopy, The University of Queensland, Brisbane, Australia.

Bio-Protocol
|March 3, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces an automated ImageJ macro to quantify cell surface ruffling, crucial for immune cell function and migration. The new method enables high-throughput analysis of ruffling changes induced by drugs or pathogens.

Keywords:
AutomationCell surfaceF-actinHigh-throuhgputImage analysisImageJ MacroMicroscopyRuffling

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Area of Science:

  • Cell Biology
  • Immunology
  • Microscopy

Background:

  • Cell surface ruffles are dynamic, F-actin-rich protrusions involved in cell migration and environmental sensing.
  • Macrophages exhibit constitutive and pathogen-induced ruffling, essential for immune surveillance and signaling.
  • Current methods for quantifying cell ruffling are manual and qualitative, limiting high-throughput analysis.

Purpose of the Study:

  • To develop an automated ImageJ macro for quantitative assessment of dorsal cell surface protrusions.
  • To provide a high-throughput screening method for analyzing changes in cell ruffling.

Main Methods:

  • Development of an automated ImageJ macro for analyzing 3D microscopy images.
  • Quantification of dorsal cell surface protrusion area and intensity.
  • Normalization of ruffling measurements for comparative analysis.

Main Results:

  • The macro automates the quantification of cell ruffling from 3D images.
  • It provides normalized measurements of ruffle area and intensity.
  • The assay is suitable for high-throughput screening applications.

Conclusions:

  • An automated ImageJ macro offers a quantitative and high-throughput solution for assessing cell ruffling.
  • This tool facilitates the study of cellular responses to stimuli like pathogens, drugs, and growth factors.
  • The developed assay aids in understanding immune cell function and migration dynamics.