Hsieh Chia-Chun, Lin Zi-Jing, Lai Lee-Jene
Experimental Division, Synchrotron Radiation Research Center, 101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu 300092, Taiwan, ROC.
J Struct Biol X. 2024 Oct 18;10:100113. doi: 10.1016/j.yjsbx.2024.100113. eCollection 2024 Dec.
Cryo-soft X-ray tomography (cryo-SXT) is a newly developed technique for imaging 3D whole cells in nearly native states. Cryo-SXT users require the preparation of numerous cryo-sample grids to use the allocated beamtime to study cellular phenomena under various conditions. Therefore, it is important to promptly prepare cryo-sample grids as efficiently and carefully as possible to minimize ice contamination on the frozen sample grid. In this study, we designed a cryo-multi-grid-box storage system, which includes a shell, funnel holder, and multi-grid-box container. Our system not only increases the number of cryo-sample grids that can be temporarily stored but also reduces the frequency of cryo grid-box container transfers, thus decreasing the probability of forming ice on the grid. We have also applied this system to A549 cryo cell grid preparation. The correlative images from cryo-light microscopy and cryo-SXT showed that limited ice had formed on the grid when preparation was performed using our system. Additionally, 3D images of mitochondria with the lamellar shape of the cristae could be observed in our cryo-SXT results. Our cryo-multi-grid-box storage system can be used for cryo-SXT and cryo-electron tomography (cryo-ET) applications.
低温软X射线断层扫描（cryo-SXT）是一种新开发的用于对近乎天然状态的三维全细胞进行成像的技术。cryo-SXT的用户需要制备大量的低温样品网格，以便利用分配的束流时间来研究各种条件下的细胞现象。因此，尽可能高效且仔细地迅速制备低温样品网格，以尽量减少冷冻样品网格上的冰污染，这一点很重要。在本研究中，我们设计了一种低温多网格盒存储系统，它包括一个外壳、漏斗支架和多网格盒容器。我们的系统不仅增加了可临时存储的低温样品网格数量，还减少了低温网格盒容器的转移频率，从而降低了网格上结冰的概率。我们还将该系统应用于A549细胞低温网格制备。低温光学显微镜和cryo-SXT的相关图像显示，使用我们的系统进行制备时，网格上形成的冰有限。此外，在我们的cryo-SXT结果中可以观察到具有板层状嵴的线粒体的三维图像。我们的低温多网格盒存储系统可用于cryo-SXT和低温电子断层扫描（cryo-ET）应用。
