CO₂-Free On-Stage Incubator for Live Cell Imaging of Cholangiocarcinoma Cell Migration on Microfluidic Device.

IF 2.3 Q3 BIOCHEMICAL RESEARCH METHODS Methods and Protocols Pub Date : 2024-09-04 DOI:10.3390/mps7050069

Shahab Ud Din, Puey Ounjai, Arthit Chairoungdua, Werasak Surareungchai

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Abstract

Long-term live cell imaging requires sophisticated and fully automated commercial-stage incubators equipped with specified inverted microscopes to regulate temperature, CO₂ content, and humidity. In this study, we present a CO₂-free on-stage incubator specifically designed for use across various cell culture platforms, enabling live cell imaging applications. A simple and transparent incubator was fabricated from acrylic sheets to be easily placed on the stages of most inverted microscopes. We successfully performed live-cell imaging of cholangiocarcinoma (CCA) cells and HeLa cell dynamics in both 2D and 3D microenvironments over three days. We also analyzed directed cell migration under high serum induction within a microfluidic device. Interesting phenomena such as "whole-colony migration", "novel type of collective cell migration" and "colony formation during cell and colony migration" are reported here for the first time, to the best of our knowledge. These phenomena may improve our understanding of the nature of cell migration and cancer metastasis.

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用于胆管癌细胞在微流控装置上迁移的活细胞成像的无二氧化碳阶段培养箱

长期活细胞成像需要配备特定倒置显微镜的精密、全自动商用阶段培养箱来调节温度、二氧化碳含量和湿度。在本研究中，我们提出了一种无二氧化碳的台上培养箱，专门设计用于各种细胞培养平台，使活细胞成像应用成为可能。我们用丙烯酸板制作了一个简单而透明的培养箱，可以方便地放置在大多数倒置显微镜的平台上。我们成功地对胆管癌（CCA）细胞和 HeLa 细胞在二维和三维微环境中的动态进行了为期三天的活细胞成像。我们还分析了微流控装置在高血清诱导下的定向细胞迁移。据我们所知，这里首次报道了 "全菌落迁移"、"新型细胞集体迁移 "和 "细胞和菌落迁移过程中的菌落形成 "等有趣的现象。这些现象可能会加深我们对细胞迁移和癌症转移本质的理解。

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