Rapid tool for cell nanoarchitecture integrity assessment.
Rapid tool for cell nanoarchitecture integrity assessment.
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DOI：
10.1016/j.jsb.2021.107801
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发表时间：
2021-12
影响因子：
3
通讯作者：
Hanein D
中科院分区：
文献类型：
--
作者：
Gaietta G;Swift MF;Volkmann N;Hanein D
随着细胞高分辨率成像技术的快速发展和普及，对结果的解释越来越依赖于周围细胞景观的三维完整性不受实验设置影响的假设。然而，唯一可用的技术，直接探测的结构完整性的全细胞制剂在纳米级是电子冷冻断层扫描，这是耗时，昂贵，复杂。我们设计了一种方便，廉价和可靠的筛选试验，以快速报告实验方案与保持纳米级全细胞制剂结构完整性的兼容性。我们的快速细胞完整性评估（RCIA）检测在室温下进行，仅依赖于光学显微镜成像。使用细胞电子冷冻断层扫描作为基准，我们验证RCIA 1准确地揭示了试剂和/或协议的不利影响，如用于病毒灭活或逮捕细胞纳米结构上的动态过程。
With the rapid increase and accessibility of high-resolution imaging technologies of cells, the interpretation of results relies more and more on the assumption that the three-dimensional integrity of the surrounding cellular landscape is not compromised by the experimental setup. However, the only available technology for directly probing the structural integrity of whole-cell preparations at the nanoscale is electron cryo-tomography, which is time-consuming, costly, and complex. We devised an accessible, inexpensive and reliable screening assay to quickly report on the compatibility of experimental protocols with preserving the structural integrity of whole-cell preparations at the nanoscale. Our Rapid Cell Integrity Assessment (RCIA) assay is executed at room temperature and relies solely on light microscopy imaging. Using cellular electron cryo-tomography as a benchmark, we verify that RCIA1 accurately unveils the adverse impact of reagents and/or protocols such as those used for virus inactivation or to arrest dynamic processes on the cellular nanoarchitecture.
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