Surface plasmon resonance imaging of excitable cells
Surface plasmon resonance imaging of excitable cells
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DOI：
10.1101/390948
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发表时间：
2018-08
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影响因子：
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通讯作者：
Carmel L. Howe;Kevin Francis Webb;Sidahmed A Abayzeed;David J. Anderson;C. Denning;Noah A. Russell
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文献类型：
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作者：
Carmel L. Howe;Kevin Francis Webb;Sidahmed A Abayzeed;David J. Anderson;C. Denning;Noah A. Russell
表面等离子体（SPs）是发生在金属/介质界面上的表面电荷密度振荡，并且对表面附近的折射率变化高度敏感。这种敏感性已被成功地用于化学和生物分析。在这些系统中，基于表面等离子体共振（SPR）的传感器检测某一点折射率的时间变化。SPR也被用于成像系统中，其中样品中折射率的空间变化提供了对比机制。SPR成像系统采用高数值孔径物镜，具有高放大倍率和近衍射极限空间分辨率。解决细胞生理学和药理学的研究问题通常需要多模态显微镜的发展，在那里可以获得互补的信息。在本文中，我们介绍了一种多模态显微镜的发展，该显微镜将SPR成像与许多其他成像方式相结合，包括明场、表观荧光、全内反射显微镜和SPR荧光显微镜。我们使用高NA物镜进行SPR和TIR显微镜，该平台已用于对活细胞培养成像，展示了荧光和无标记技术。SPR和TIR成像系统都具有宽视场（~300 μ m），允许从多个细胞进行测量，同时保持足够的分辨率来成像精细的细胞过程。通过对干细胞来源的心肌细胞收缩的空间变化进行成像，证明了该平台对活细胞进行无标记功能成像的能力。这项技术显示了在发育过程中很长一段时间内对培养细胞进行无创成像的希望。
Surface plasmons (SPs) are surface charge density oscillations occuring at a metal/dieletric interface and are highly sensitive to refractive index variations adjacent to the surface. This sensitivity has been exploited successfully for chemical and biological assays. In these systems, a surface plasmon resonance (SPR)-based sensor detects temporal variations in the refractive index at a point. SPR has also been used in imaging systems where the spatial variations of refractive index in the sample provide the contrast mechanism. SPR imaging systems using high numerical aperture (NA) objective lenses have been designed to image adherent live cells with high magnification and near-diffraction limited spatial resolution. Addressing research questions in cell physiology and pharmacology often requires the development of a multimodal microscope where complementary information can be obtained. In this paper, we present the development of a multimodal microscope that combines SPR imaging with a number of additional imaging modalities including bright-field, epifluorescence, total internal reflection microscopy and SPR fluorescence microscopy. We used a high NA objective lens for SPR and TIR microscopy and the platform has been used to image live cell cultures demonstrating both fluorescent and label-free techniques. Both the SPR and TIR imaging systems feature a wide field of view (~300 µm) that allows measurements from multiple cells whilst maintaining a resolution sufficient to image fine cellular processes. The capability of the platform to perform label-free functional imaging of living cells was demonstrated by imaging the spatial variations in contractions from stem cell-derived cardiomyocytes. This technique shows promise for non-invasive imaging of cultured cells over very long periods of time during development.