Developing Technologies for Correlative Cryo-Imaging Pipelines.
Developing Technologies for Correlative Cryo-Imaging Pipelines.
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开发相关冷冻成像管道技术。
DOI：
10.1093/micmic/ozad067.519
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Cai,Kai
中科院分区：
文献类型：
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作者：
Wright,ElizabethR;Yang,Jae;Sibert,Bryan;Larson,Matthew;Kim,JosephY;Parrell,Daniel;Sanchez,JuanC;Kumar,Anil;Cai,Kai
细胞在所有活着的有机体中无处不在。细胞要么在它们的奇点上是“简单的”，要么是复杂的多细胞群落的“积木”。显微镜技术的许多进步，特别是冷冻显微镜技术，使研究人员能够开始研究这些众所周知的“酶袋”中的细胞、细胞器和大分子的有限细节。低温电子断层扫描(Cryo-ET)与其他成像手段相结合，包括荧光显微镜(FLM)和聚焦离子束扫描电子显微镜(FIB-SEM)，支持了细胞生物学的新的复兴[1]。然而，这些技术仍处于初级阶段，工作流程、软件和硬件正在经历快速发展，以改进标本保存、成像以及数据处理和分析。在这篇演讲中，我们将描述一些正在进行的开发，以通过衬底微图案化[2]和替代冷冻方法来改进样品保存；使用相关器[3]进行图像和位置关联；使用蒙太奇并行阵列低温层析成像(MPACT)采集方案来获取大范围样本的电子断层扫描数据[4]；以及用于三维(3D)数据处理和分析的包装器和管道。
Cells are ubiquitous to all living organisms. Cells are either ‘simple’in their singularity or are the ‘building blocks’ of complex multi-cellular communities. Many advances in microscopy, especially cryo-microscopy techniques, have allowed researchers to begin to investigate the finite detail of cells, organelles, and macromolecules within these proverbial ‘bags of enzymes.’Cryo-electron tomography (cryo-ET) combined with other imaging modalities including fluorescence light microscopy (FLM) and focused ion beam-scanning electron microscopy (FIB-SEM) have supported a new renaissance in cell biology [1]. However, these technologies are still in their infancy and workflows, software, and hardware are undergoing rapid developments to improve specimen preservation, imaging, and data processing and analysis. In this presentation, we will describe some of the developments underway for improving sample preservation via substrate micropatterning [2] and alternative freezing approaches; image and position correlation using CorRelator [3]; using Montage Parallel Array Cryo-Tomography (MPACT) collection schemes to acquire electron tomography data over large regions of a specimen [4]; and wrappers and pipelines for three-dimensional (3D) data processing and analysis.