<br>悬浮贴壁 3D 细胞培养物的高通量 3D 成像流式细胞术,Small Methods

当前位置： X-MOL 学术 › Small Methods › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

悬浮贴壁 3D 细胞培养物的高通量 3D 成像流式细胞术
Small Methods ( IF 10.7 ) Pub Date : 2023-12-22 , DOI: 10.1002/smtd.202301318
Minato Yamashita ₁ , Miu Tamamitsu ₁ , Hiromi Kirisako ₁ , Yuki Goda ₁ , Xiaoyao Chen ₁ , Kazuki Hattori ₁ , Sadao Ota ₁

Affiliation

3D 细胞培养对于重现体内环境是不可或缺的。在众多 3D 培养方法中，在水凝胶珠上培养贴壁细胞以形成球状结构是在贴壁状态下维持高细胞活力和功能的强大策略。然而，缺乏对水凝胶支架上培养的单个细胞进行 3D 成像的高通量、可扩展技术。本研究报告了用于分析球体模型的高通量、可扩展 3D 成像流式细胞术平台的开发。该平台是通过将单物镜荧光光片显微镜与结合了流体动力学和声流控聚焦技术的微流体装置集成来实现的。这种集成实现了前所未有的高通量和可扩展的光流控 3D 成像，可处理由 28 117 个细胞 min ^-1组成的 1310 个球体。获得的大数据集能够对粘附细胞和悬浮细胞的核形态进行精确量化和比较，揭示粘附细胞具有较小的细胞核和较小的圆形表面。该平台在分析 3D 培养模型中亚细胞结构形态方面具有高通量、稳健性和精确度，在各种生物医学分析中具有广阔的前景，包括基于图像的球体或类器官药物表型筛选。

"点击查看英文标题和摘要"

High-Throughput 3D Imaging Flow Cytometry of Suspended Adherent 3D Cell Cultures

3D cell cultures are indispensable in recapitulating in vivo environments. Among the many 3D culture methods, culturing adherent cells on hydrogel beads to form spheroid-like structures is a powerful strategy for maintaining high cell viability and functions in the adherent states. However, high-throughput, scalable technologies for 3D imaging of individual cells cultured on the hydrogel scaffolds are lacking. This study reports the development of a high throughput, scalable 3D imaging flow cytometry platform for analyzing spheroid models. This platform is realized by integrating a single objective fluorescence light-sheet microscopy with a microfluidic device that combines hydrodynamic and acoustofluidic focusing techniques. This integration enabled unprecedentedly high-throughput and scalable optofluidic 3D imaging, processing 1310 spheroids consisting of 28 117 cells min⁻¹. The large dataset obtained enables precise quantification and comparison of the nuclear morphology of adhering and suspended cells, revealing that the adhering cells have smaller nuclei with less rounded surfaces. This platform's high throughput, robustness, and precision for analyzing the morphology of subcellular structures in 3D culture models hold promising potential for various biomedical analyses, including image-based phenotypic screening of drugs with spheroids or organoids.

更新日期：2023-12-22

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文