Live-Cell F&#246;rster Resonance Energy Transfer Imaging of Metabolically Regulated Akt Activation Dynamics in HepG2 Cells

Javed Akhtar; Muhammad Imran; Jiahe Wang; Gang Ma; Guanyu Wang

doi:10.3791/68075

HepG2 细胞中代谢调节的 Akt 活化动力学的活细胞 Förster 共振能量转移成像

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Cancer Research

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JoVE Journal Cancer Research

Live-Cell Förster Resonance Energy Transfer Imaging of Metabolically Regulated Akt Activation Dynamics in HepG2 Cells

HepG2 细胞中代谢调节的 Akt 活化动力学的活细胞 Förster 共振能量转移成像

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08:03 min

May 23, 2025

DOI: 10.3791/68075-v

Javed Akhtar^1,2,3,4,5, Muhammad Imran⁶, Jiahe Wang^1,2,3, Gang Ma⁵, Guanyu Wang^1,2,3

¹Laboratory of Biocomplexity and Engineering Biology, School of Medicine,The Chinese University of Hong Kong, ²Futian Biomedical Innovation R&D Center,The Chinese University of Hong Kong, ³Ciechanover Institute of Precision and Regenerative Medicine, School of Medicine,The Chinese University of Hong Kong, ⁴Department of Immunology and Microbiology, School of Life Sciences,Southern University of Science and Technology, ⁵Department of Biology, School of Life Sciences,Southern University of Science and Technology, ⁶Department of Computer Science and IT,University of Southern Punjab

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在这里，我们提出了一种方案来量化活 HepG2 细胞中 Akt 激活和磷酸化的时空动力学。Förster 共振能量转移（FRET）成像是一种强大的工具，可为癌细胞中的胰岛素信号通路和代谢调节提供有价值的见解。

细胞异质性是准确描述细胞动力学复杂性的障碍，这些动力学基本上是平均的。这就是我们在这里对单个细胞进行成像的原因。基因编码的 FRET 生物传感器的最新发展提高了特异性，并彻底改变了细胞内 Akt 的实时跟踪，这有助于理解异基因和开发个性化药物疗法。

癌症和糖尿病是通过复杂的代谢途径进行的。为了解码癌症中胰岛素信号通路的生物复杂性，我们使用了单细胞分辨率的尖端活细胞 FRET成像。我们发现，优化共聚焦显微镜、硬件和软件配置以及微调激光功率和调整探测器增益对于最大限度地减少FRET信号中的任何噪声和光谱串扰至关重要。

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