Correlative super-resolution fluorescence and electron microscopy using conventional fluorescent proteins in vacuo.
Correlative super-resolution fluorescence and electron microscopy using conventional fluorescent proteins in vacuo.
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DOI：
10.1016/j.jsb.2017.05.013
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发表时间：
2017-08
影响因子：
3
通讯作者：
Collinson LM
中科院分区：
文献类型：
--
作者：
Peddie CJ;Domart MC;Snetkov X;O'Toole P;Larijani B;Way M;Cox S;Collinson LM
超分辨率光学显微镜、相关光学和电子显微镜以及体积电子显微镜正在彻底改变生物样品的检测和理解方式。在这里，我们联合收割机这些方法提供超准确的荧光蛋白的相关性，细胞结构。我们表明，YFP和GFP具有增强的闪烁特性时，嵌入在丙烯酸树脂和成像在部分真空下，使在真空中单分子定位显微镜。在传统的基于切片的相关显微镜实验中，样品必须在成像系统之间移动和/或进一步操作以获得最佳观察效果。这些步骤可能在样本中引入不期望的改变，并且使成像模态之间的相关性复杂化。我们通过使用集成光学显微镜的扫描电子显微镜来获取定位和电子显微镜图像，然后可以精确地关联，从而避免了这些问题。收集的数据，从显微镜切片也提高了轴向分辨率和信噪比的原始本地化显微镜数据。将数据收集扩展到一系列部分，将允许在前所未有的数量上进行三维关联。该技术的性能在牛痘病毒（具有YFP）和细胞膜中的二酰基甘油（具有GFP）上得到证明。
Super-resolution light microscopy, correlative light and electron microscopy, and volume electron microscopy are revolutionising the way in which biological samples are examined and understood. Here, we combine these approaches to deliver super-accurate correlation of fluorescent proteins to cellular structures. We show that YFP and GFP have enhanced blinking properties when embedded in acrylic resin and imaged under partial vacuum, enabling in vacuo single molecule localisation microscopy. In conventional section-based correlative microscopy experiments, the specimen must be moved between imaging systems and/or further manipulated for optimal viewing. These steps can introduce undesirable alterations in the specimen, and complicate correlation between imaging modalities. We avoided these issues by using a scanning electron microscope with integrated optical microscope to acquire both localisation and electron microscopy images, which could then be precisely correlated. Collecting data from ultrathin sections also improved the axial resolution and signal-to-noise ratio of the raw localisation microscopy data. Expanding data collection across an array of sections will allow 3-dimensional correlation over unprecedented volumes. The performance of this technique is demonstrated on vaccinia virus (with YFP) and diacylglycerol in cellular membranes (with GFP).
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影响因子：
4.6
作者：
Johnson E;Seiradake E;Jones EY;Davis I;Grünewald K;Kaufmann R
通讯作者：
Kaufmann R
影响因子：
64.8
作者：
Dickson, RM;Cubitt, AB;Moerner, WE
通讯作者：
Moerner, WE
DOI：
10.1007/s12154-015-0143-3
发表时间：
2015-10-01
期刊：
Journal of Chemical Biology
影响因子：
--
作者：
Brama, Elisabeth;Peddie, Christopher J.;Collinson, Lucy M.
通讯作者：
Collinson, Lucy M.
影响因子：
3.7
作者：
Domart MC;Hobday TM;Peddie CJ;Chung GH;Wang A;Yeh K;Jethwa N;Zhang Q;Wakelam MJ;Woscholski R;Byrne RD;Collinson LM;Poccia DL;Larijani B
通讯作者：
Larijani B
DOI：
10.1073/pnas.0700059104
发表时间：
2007-04-17
影响因子：
11.1
作者：
Henderson, J. Nathan;Ai, Hui-wang;Remington, S. James
通讯作者：
Remington, S. James