CorRelator: Interactive software for real-time high precision cryo-correlative light and electron microscopy.
CorRelator: Interactive software for real-time high precision cryo-correlative light and electron microscopy.
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DOI：
10.1016/j.jsb.2021.107709
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发表时间：
2021-06
影响因子：
3
通讯作者：
Wright ER
中科院分区：
文献类型：
--
作者：
Yang JE;Larson MR;Sibert BS;Shrum S;Wright ER
冷冻相关光电子显微镜（CLEM）是一种利用荧光显微镜（FLM）的时空线索，通过冷冻电子显微镜（cryo-EM）研究生物样品高分辨率超微结构的技术。Cryo-CLEM为从细胞环境中识别和区分荧光标记的蛋白质、大分子复合物和细胞器提供了优势。如何在不同的显微镜平台上实施相关工作流程和软件工具以支持自动化冷冻EM数据采集仍然存在挑战。在这里，我们介绍了CorRelator：一个开源的桌面应用程序，在cryo-FLM和实时cryo-EM/ET自动数据收集之间架起了桥梁。CorRelator实现了像素坐标到载物台位置的转换，可实现灵活、高精度的实时和采集后相关。CorRelator可以集成到cryo-CLEM工作流程中，并轻松适应标准荧光和透射电子显微镜（TEM）系统配置。使用几种FLM和TEM仪器在活细胞和低温条件下对CorRelator进行了基准测试，表明CorRelator通过软件辅助和交互式对齐的组合可靠地支持实时，自动相关的cryo-EM/ET采集。CorRelator是一个跨平台的软件包，具有直观的图形用户界面（GUI），可指导用户完成相关过程。CorRelator源代码可在https://github.com/wright-cemrc-projects/corr上获得。
Cryo-correlative light and electron microscopy (CLEM) is a technique that uses the spatiotemporal cues from fluorescence light microscopy (FLM) to investigate the high-resolution ultrastructure of biological samples by cryo-electron microscopy (cryo-EM). Cryo-CLEM provides advantages for identifying and distinguishing fluorescently labeled proteins, macromolecular complexes, and organelles from the cellular environment. Challenges remain on how correlation workflows and software tools are implemented on different microscope platforms to support automated cryo-EM data acquisition. Here, we present CorRelator: an open-source desktop application that bridges between cryo-FLM and real-time cryo-EM/ET automated data collection. CorRelator implements a pixel-coordinate-to-stage-position transformation for flexible, high accuracy on-the-fly and post-acquisition correlation. CorRelator can be integrated into cryo-CLEM workflows and easily adapted to standard fluorescence and transmission electron microscope (TEM) system configurations. CorRelator was benchmarked under live-cell and cryogenic conditions using several FLM and TEM instruments, demonstrating that CorRelator reliably supports real-time, automated correlative cryo-EM/ET acquisition, through a combination of software-aided and interactive alignment. CorRelator is a cross-platform software package featuring an intuitive Graphical User Interface (GUI) that guides the user through the correlation process. CorRelator source code is available at: https://github.com/wright-cemrc-projects/corr.
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