2025/04/21 by Florian Aubermann, Senne Seneca, Tomáš Hofman +6 · 1 voice · 4 citations
Biochemistry, Genetics and Molecular Biology · Engineering · Immunology and Microbiology · #Biology #Cell #Cell biology #Computational biology #Computer science #Cytotoxicity #Effector #Genetics #Immune Cell Function and Interaction #In vitro #Innovative Microfluidic and Catalytic Techniques Innovation #Materials science #Microfluidics #Modular design #Nanotechnology #Natural killer cell #Phenotype #Single-cell and spatial transcriptomics
paper · pdf · doi:10.1002/smtd.202500236
published in Small Methods 9(8), e2500236 (Wiley)
openalex publication_date 2025/04/21 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/06
Natural Killer (NK) cells, as key effector cells of the innate immune system, display high heterogeneity in their ability to kill target cells. The underlying mechanisms remain poorly understood. Here, a droplet-based microfluidic platform is presented to identify and select NK cells with serial killing ability. To this end, primary human NK cells are encapsulated with several target cells using an efficient negative pressure-based droplet generator. Capitalizing on the large number of possible killing events due to quantization into droplets, a convolutional neural network analysis pipeline is developed to quantify the cytotoxicity and abundance of serial killing events with high accuracy. To physically select NK cells based on their serial killing ability, MultiCell-Sort - an advanced real-time image-based droplet sorting module - is presented. While conventional single-cell sorters mostly evaluate intrinsically-encoded properties, such as protein expression levels, MultiCell-Sort can select living NK cells based on complex functional phenotypes emerging from multiple cell-cell interactions within the droplet. This novel microfluidic phenotyping platform hereby allows the potential integration of complementary techniques to provide an understanding of regulators and markers underlying the heterogeneous nature of NK cell functional phenotypes.