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Related Experiment Video

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Modular Droplet-Based Microfluidic Platform for Functional Phenotypic Screening of Natural Killer Cells.

Florian Aubermann1,2,3, Senne Seneca1,2, Tomáš Hofman4

  • 1Department of Cellular Biophysics, Max Planck Institute for Medical Research, 69120, Heidelberg, Germany.

Small Methods
|April 21, 2025
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Summary

This study introduces a microfluidic platform to analyze Natural Killer (NK) cell serial killing ability. The technology enables the selection of NK cells based on their functional killing phenotypes, advancing innate immunity research.

Keywords:
droplet‐based microfluidicsnatural killer cell phenotypesnegative pressuresupervised deep learning

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Area of Science:

  • Immunology
  • Microfluidics
  • Cell Biology

Background:

  • Natural Killer (NK) cells are crucial for innate immunity but show variable target cell killing abilities.
  • The mechanisms driving NK cell functional heterogeneity are not well understood.

Purpose of the Study:

  • To develop a droplet-based microfluidic platform for identifying and selecting NK cells with serial killing capacity.
  • To enable the study of complex functional phenotypes in NK cells.

Main Methods:

  • Encapsulation of primary human NK cells with target cells in droplets using a microfluidic device.
  • Development of a convolutional neural network (CNN) pipeline for quantifying cytotoxicity and serial killing events.
  • Implementation of MultiCell-Sort, a real-time image-based droplet sorter, for functional cell selection.

Main Results:

  • The platform accurately quantifies NK cell cytotoxicity and serial killing events.
  • MultiCell-Sort successfully selects live NK cells based on functional phenotypes derived from cell-cell interactions.
  • Demonstration of a novel microfluidic approach for phenotyping heterogeneous NK cell populations.

Conclusions:

  • The presented microfluidic platform offers a novel method for analyzing and sorting NK cells based on their functional killing capabilities.
  • This technology can facilitate the discovery of regulators and markers associated with NK cell heterogeneity.
  • Advances understanding of innate immune effector cell function and selection.