具有受激发射拉曼散射显微镜的延时活细胞成像的柔性腔。
A Flexible Chamber for Time-Lapse Live-Cell Imaging with Stimulated Raman Scattering Microscopy.
机构信息
Department of Biomedical Engineering, Binghamton University, State University of New York.
Department of Biomedical Engineering, Binghamton University, State University of New York;
出版信息
J Vis Exp. 2022 Aug 31(186). doi: 10.3791/64449.
Stimulated Raman scattering (SRS) microscopy is a label-free chemical imaging technology. Live-cell imaging with SRS has been demonstrated for many biological and biomedical applications. However, long-term time-lapse SRS imaging of live cells has not been widely adopted. SRS microscopy often uses a high numerical aperture (NA) water-immersion objective and a high NA oil-immersion condenser to achieve high-resolution imaging. In this case, the gap between the objective and the condenser is only a few millimeters. Therefore, most commercial stage-top environmental chambers cannot be used for SRS imaging because of their large thickness with a rigid glass cover. This paper describes the design and fabrication of a flexible chamber that can be used for time-lapse live-cell imaging with transmitted SRS signal detection on an upright microscope frame. The flexibility of the chamber is achieved by using a soft material - a thin natural rubber film. The new enclosure and chamber design can be easily added to an existing SRS imaging setup. The testing and preliminary results demonstrate that the flexible chamber system enables stable, long-term, time-lapse SRS imaging of live cells, which can be used for various bioimaging applications in the future.
受激拉曼散射（SRS）显微镜是一种无标记的化学成像技术。已经证明，SRS 可用于许多生物和生物医学应用的活细胞成像。然而，长期的活细胞 SRS 成像并未被广泛采用。SRS 显微镜通常使用高数值孔径（NA）水浸物镜和高 NA 油浸聚光镜来实现高分辨率成像。在这种情况下，物镜和聚光镜之间的间隙只有几毫米。因此，由于其刚性玻璃盖的厚度较大，大多数商用的台式环境室都不能用于 SRS 成像。本文介绍了一种柔性室的设计和制造，该柔性室可用于在正置显微镜架上进行具有透射 SRS 信号检测的活细胞延时成像。通过使用柔软的材料-薄天然橡胶膜来实现腔室的灵活性。新的外壳和腔室设计可以很容易地添加到现有的 SRS 成像设置中。测试和初步结果表明，柔性腔系统能够稳定、长期、延时地对活细胞进行 SRS 成像，这可用于未来的各种生物成像应用。
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