Smith Elizabeth A, Cinquin Bertrand P, Do Myan, McDermott Gerry, Le Gros Mark A, Larabell Carolyn A
Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; Physical BioSciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
Ultramicroscopy. 2014 Aug;143:33-40. doi: 10.1016/j.ultramic.2013.10.013. Epub 2013 Nov 7.
Correlated imaging is the process of imaging a specimen with two complementary modalities, and then combining the two data sets to create a highly informative, composite view. A recent implementation of this concept has been the combination of soft x-ray tomography (SXT) with fluorescence cryogenic microscopy (FCM). SXT-FCM is used to visualize cells that are held in a near-native, cryopreserved. The resultant images are, therefore, highly representative of both the cellular architecture and molecular organization in vivo. SXT quantitatively visualizes the cell and sub-cellular structures; FCM images the spatial distribution of fluorescently labeled molecules. Here, we review the characteristics of SXT-FCM, and briefly discuss how this method compares with existing correlative imaging techniques. We also describe how the incorporation of a cryo-rotation stage into a cryogenic fluorescence microscope allows acquisition of fluorescence cryogenic tomography (FCT) data. FCT is optimally suited for correlation with SXT, since both techniques image the specimen in 3-D, potentially with similar, isotropic spatial resolution.
关联成像就是用两种互补模态对样本进行成像，然后将这两个数据集合并，以创建一个信息丰富的合成视图的过程。该概念最近的一个应用实例是软X射线断层扫描（SXT）与荧光低温显微镜（FCM）的结合。SXT-FCM用于可视化保存在接近天然状态的冷冻保存细胞。因此，所得图像高度代表了体内的细胞结构和分子组织。SXT定量地可视化细胞和亚细胞结构；FCM对荧光标记分子的空间分布进行成像。在这里，我们回顾了SXT-FCM的特点，并简要讨论了该方法与现有关联成像技术的比较。我们还描述了如何将低温旋转台整合到低温荧光显微镜中以获取荧光低温断层扫描（FCT）数据。FCT最适合与SXT进行关联，因为这两种技术都对样本进行三维成像，潜在空间分辨率可能相似且各向同性。
