A Thermoplastic Chip for 2D and 3D Correlative Assays Combining Screening and High-resolution Imaging of Immune Cell Responses

Overview

Journal Cell Rep Methods

Date 2025 Jan 18

PMID 39826552

Authors

Hanna van Ooijen

Quentin Verron

Hanqing Zhang

Patrick A Sandoz

Thomas W Frisk

Valentina Carannante

Karl Olofsson

Arnika K Wagner

Niklas Sandstrom

Bjorn Onfelt

Affiliations

Soon will be listed here.

Abstract

We present an easy-to-use, disposable, thermoplastic microwell chip designed to support screening and high-resolution imaging of single-cell behavior in two- and three-dimensional (2D and 3D) cell cultures. We show that the chip has excellent optical properties and provide simple protocols for efficient long-term cell culture of suspension and adherent cells, the latter grown either as monolayers or as hundreds of single, uniformly sized spheroids. We then demonstrate the applicability of the system for single-cell analysis by correlating the dynamic cytotoxic response of single immune cells grown under different metabolic conditions to their intracellular cytolytic load at the end of the assay. Additionally, we illustrate highly multiplex cytotoxicity screening of tumor spheroids in the chip, comparing the effect of environment cues characteristic of the tumor microenvironment on natural killer (NK)-cell-induced killing. Following the functional screening, we perform high-resolution 3D immunofluorescent imaging of infiltrating NK cells within the spheroid volumes.

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