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Author:
Date:
2016-01-05
[Abstract] Light microscopy is the standard tool for studying sub-cellular structures however, owing to the diffractive properties of light, resolution is limited to 200 nm. Super-resolution microscopy methods circumvent this limit, offering greater resolution, particularly when studying fluorescently labeled sub-cellular structures. Super-resolution methods such as 3D-SIM (Structured Illumination Microscopy) fill a useful niche between confocal and electron microscopy. We have previously had success using fixed plant tissue samples with 3D-SIM (Bell and Oparka, 2014). However, sensitive structures can be altered by fixation and embedding procedures, so we developed a method for imaging live cells. In this protocol we used 3D-SIM to image the ER and Hechtian Strands in live, plasmolysed BY2 cells.
[摘要] 光学显微镜是用于研究亚细胞结构的标准工具，然而，由于光的衍射性质，分辨率限于200nm。 超分辨率显微镜方法规避这一限制，提供更大的分辨率，特别是当研究荧光标记的亚细胞结构。 超分辨率方法，如3D-SIM（结构照明显微镜）填充共聚焦和电子显微镜之间有用的利基。 我们以前已经成功使用固定植物组织样本与3D-SIM（贝尔和Oparka，2014）。 然而，敏感结构可以通过固定和嵌入程序改变，所以我们开发了一种成像活细胞的方法。 在这个协议中，我们使用3D-SIM成像ER和Hechtian链在活的，plasmolysed BY2细胞。
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