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Related Concept Videos

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Related Experiment Video

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Quantitative Microscopy for Cell-Surface and Cell-Cell Interactions in Immunology.

Beatriz Díaz-Bello1, Dalia El Arawi1, Rémy Torro1,2

  • 1Aix-Marseille University, CNRS, INSERM, LAI, Turing Center for Living Systems, Marseille, France.

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|September 15, 2025
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Summary

This study introduces novel optical microscopy assays for analyzing Natural Killer (NK) cell interactions with cancer cells and surfaces. These methods enable precise quantification of cell-cell cytotoxicity and receptor-ligand dynamics for immunotherapy research.

Keywords:
ADCCCell spreadingFluorescence microscopyImage analysis softwareImmunotherapyRICMSingle-cell dynamicsTime-lapse microscopy

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Area of Science:

  • * Immunology
  • * Cell Biology
  • * Optical Microscopy

Background:

  • * Cell-surface and cell-cell interactions are crucial for understanding receptor-ligand dynamics, immune responses, and therapeutic applications like cancer immunotherapy.
  • * Traditional methods often rely on ensemble measurements or indirect readouts, limiting detailed analysis of cellular behavior.
  • * Optical microscopy offers direct, time-lapsed, cell-level observation, providing advantages over ensemble or indirect techniques.

Purpose of the Study:

  • * To describe two complementary microscopy-based assays for evaluating Natural Killer (NK) cell interactions.
  • * To quantify dynamic cell-surface ligand binding between primary NK cells and a cancer-mimicking surface.
  • * To assess antibody-dependent cell-mediated cytotoxicity (ADCC) between NK cells and tumor cells using advanced imaging and analysis.

Main Methods:

  • * Development of a cell-surface ligand binding assay using label-free imaging to study NK cell spreading on antibody-coated surfaces.
  • * Implementation of a cell-cell interaction assay for evaluating ADCC via fluorescence imaging and deep learning-based death detection.
  • * Utilization of Celldetective, an open-source GUI, for quantitative analysis of cell interaction dynamics from 2D time-lapse microscopy data.

Main Results:

  • * Enabled quantitative analysis of dynamic NK cell spreading on antibody-coated surfaces at varying antibody concentrations.
  • * Provided high-resolution evaluation of NK cell-mediated ADCC against tumor cells.
  • * Facilitated direct observation and quantification of cellular morphology, motility, and interactions at single-cell resolution.

Conclusions:

  • * The described microscopy assays offer a powerful, adaptable approach for studying cell interactions in immunology and beyond.
  • * These methods, coupled with open-source software, facilitate detailed mechanistic analysis of immune cell functions, including cytotoxicity and synapse formation.
  • * The protocol supports antibody evaluation and therapeutic development, particularly in cancer immunotherapy, by providing precise, quantitative insights into cellular dynamics.