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一种用于二维和三维相关分析的热塑性芯片，可结合免疫细胞反应的筛选和高分辨率成像。

A thermoplastic chip for 2D and 3D correlative assays combining screening and high-resolution imaging of immune cell responses.

作者信息

van Ooijen Hanna, Verron Quentin, Zhang Hanqing, Sandoz Patrick A, Frisk Thomas W, Carannante Valentina, Olofsson Karl, Wagner Arnika K, Sandström Niklas, Önfelt Björn

机构信息

Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden.

Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.

出版信息

Cell Rep Methods. 2025 Jan 27;5(1):100965. doi: 10.1016/j.crmeth.2025.100965. Epub 2025 Jan 17.

DOI:10.1016/j.crmeth.2025.100965
PMID:39826552
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11841093/
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.

摘要

我们展示了一种易于使用的一次性热塑性微孔芯片，其设计用于支持二维和三维（2D和3D）细胞培养中单细胞行为的筛选和高分辨率成像。我们表明该芯片具有出色的光学特性，并提供了简单的方案，用于悬浮细胞和贴壁细胞的高效长期培养，后者可以单层生长或形成数百个大小均匀的单个球体。然后，我们通过将在不同代谢条件下生长的单个免疫细胞的动态细胞毒性反应与其在试验结束时的细胞内溶细胞负荷相关联，证明了该系统在单细胞分析中的适用性。此外，我们展示了芯片中肿瘤球体的高度多重细胞毒性筛选，比较了肿瘤微环境特征性环境线索对自然杀伤（NK）细胞诱导杀伤的影响。在功能筛选之后，我们对球体体积内浸润的NK细胞进行了高分辨率3D免疫荧光成像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/18d051c7f0d7/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/762e4a02f6dc/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/7ebf9f50c87f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/374b39329a8e/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/19fa1b430055/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/474279a82cae/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/e25b75591da1/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/18d051c7f0d7/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/762e4a02f6dc/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/7ebf9f50c87f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/374b39329a8e/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/19fa1b430055/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/474279a82cae/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/e25b75591da1/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d358/11841093/18d051c7f0d7/gr6.jpg

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